Rui:TotoRNAseq protocol: Difference between revisions
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>RuiLiu |
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==totoRNA library structure== | ==totoRNA library structure== | ||
===Standard primers=== | |||
[[File:Libraries_ILMN.jpg]] | [[File:Libraries_ILMN.jpg]] | ||
[[File:Libraries_totoRNA-5.jpg | ===totoRNAseq primers=== | ||
[[File:Libraries_totoRNA-5.jpg]] | |||
[[File:Libraries_totoRNA-3.jpg]] | |||
==totoRNA primer sequence== | |||
* The 5' adaptors list | |||
{| {{table}} border=1 | |||
| align="center" style="background:#f0f0f0;"|'''Tube ID''' | |||
| align="center" style="background:#f0f0f0;"|'''Primer full name''' | |||
| align="center" style="background:#f0f0f0;"|'''Primer full sequence''' | |||
| align="center" style="background:#f0f0f0;"|'''PE_F-N10''' | |||
| align="center" style="background:#f0f0f0;"|'''Barcode''' | |||
| align="center" style="background:#f0f0f0;"|'''TSO''' | |||
|- | |||
| TSO.r01||BR_TSO_N10_BC6||ACACTCTTTCCCTACACGACGCTCTTCCGATCTNNNdUNNNNdUNNNTTTAGGrGrGrG||ACACTCTTTCCCTACACGACGCTCTTCCGATCTNNN/ideoxyU/NNNN/ideoxyU/NNN||TTTAGG||rGrGrG | |||
|- | |||
| TSO.r02||B_TSO_N10||ACACTCTTTCCCTACACGACGNNNUNNNNUNNNrGrGrG||ACACTCTTTCCCTACACGACGNNN/ideoxyU/NNNN/ideoxyU/NNN||\||rGrGrG | |||
|- | |||
| TSO.r03||B.N10.TSO.NH2||ACACTCTTTCCCTACACGACGUNNNUNNNNUNNNrGrGrG/3AmMO/||ACACTCTTTCCCTACACGACGNNN/ideoxyU/NNNN/ideoxyU/NNN||\||rGrGrG | |||
|- | |||
| TSO.id01||B.TSO.N10.BC8.Idx1||ACACTCTTTCCCTACACGACG/ideoxyU//ideoxyU/NNNNNNNNNNTCTGAGACrGrGrG||ACACTCTTTCCCTACACGACG/ideoxyU//ideoxyU/NNNNNNNNNN||TCTGAGAC||rGrGrG | |||
|- | |||
| TSO.id02||B.TSO.N10.BC8.Idx2||ACACTCTTTCCCTACACGACG/ideoxyU//ideoxyU/NNNNNNNNNNTGCCCAAGrGrGrG||ACACTCTTTCCCTACACGACG/ideoxyU//ideoxyU/NNNNNNNNNN||TGCCCAAG||rGrGrG | |||
|- | |||
| TSO.id03||B.TSO.N10.BC8.Idx3||ACACTCTTTCCCTACACGACG/ideoxyU//ideoxyU/NNNNNNNNNNGACTCGCTrGrGrG||ACACTCTTTCCCTACACGACG/ideoxyU//ideoxyU/NNNNNNNNNN||GACTCGCT||rGrGrG | |||
|- | |||
| TSO.id04||B.TSO.N10.BC8.Idx4||ACACTCTTTCCCTACACGACG/ideoxyU//ideoxyU/NNNNNNNNNNCCACTCCArGrGrG||ACACTCTTTCCCTACACGACG/ideoxyU//ideoxyU/NNNNNNNNNN||CCACTCCA||rGrGrG | |||
|- | |||
| TSO.id05||B.TSO.N10.BC8.Idx5||ACACTCTTTCCCTACACGACG/ideoxyU//ideoxyU/NNNNNNNNNNGAACAGGGrGrGrG||ACACTCTTTCCCTACACGACG/ideoxyU//ideoxyU/NNNNNNNNNN||GAACAGGG||rGrGrG | |||
|- | |||
| TSO.id06||B.TSO.N10.BC8.Idx6||ACACTCTTTCCCTACACGACG/ideoxyU//ideoxyU/NNNNNNNNNNCTGCTGATrGrGrG||ACACTCTTTCCCTACACGACG/ideoxyU//ideoxyU/NNNNNNNNNN||CTGCTGAT||rGrGrG | |||
|- | |||
| TSO.id07||B.TSO.N10.BC8.Idx7||ACACTCTTTCCCTACACGACG/ideoxyU//ideoxyU/NNNNNNNNNNCTAGTATCrGrGrG||ACACTCTTTCCCTACACGACG/ideoxyU//ideoxyU/NNNNNNNNNN||CTAGTATC||rGrGrG | |||
|- | |||
| TSO.id08||B.TSO.N10.BC8.Idx8||ACACTCTTTCCCTACACGACG/ideoxyU//ideoxyU/NNNNNNNNNNGGCTGAAArGrGrG||ACACTCTTTCCCTACACGACG/ideoxyU//ideoxyU/NNNNNNNNNN||GGCTGAAA||rGrGrG | |||
|- | |||
| TSO.id09||B.TSO.N10.BC8.Idx9||ACACTCTTTCCCTACACGACG/ideoxyU//ideoxyU/NNNNNNNNNNGAGTGCTTrGrGrG||ACACTCTTTCCCTACACGACG/ideoxyU//ideoxyU/NNNNNNNNNN||GAGTGCTT||rGrGrG | |||
|- | |||
| TSO.id10||B.TSO.N10.BC8.Idx10||ACACTCTTTCCCTACACGACG/ideoxyU//ideoxyU/NNNNNNNNNNATACGCCArGrGrG||ACACTCTTTCCCTACACGACG/ideoxyU//ideoxyU/NNNNNNNNNN||ATACGCCA||rGrGrG | |||
|- | |||
| TSO.id11||B.TSO.N10.BC8.Idx11||ACACTCTTTCCCTACACGACG/ideoxyU//ideoxyU/NNNNNNNNNNATGCGTTArGrGrG||ACACTCTTTCCCTACACGACG/ideoxyU//ideoxyU/NNNNNNNNNN||ATGCGTTA||rGrGrG | |||
|- | |||
| TSO.id12||B.TSO.N10.BC8.Idx12||ACACTCTTTCCCTACACGACG/ideoxyU//ideoxyU/NNNNNNNNNNTCACCCTTrGrGrG||ACACTCTTTCCCTACACGACG/ideoxyU//ideoxyU/NNNNNNNNNN||TCACCCTT||rGrGrG | |||
|- | |||
| TSO.id13||B.TSO.N10.BC8.Idx13||ACACTCTTTCCCTACACGACG/ideoxyU//ideoxyU/NNNNNNNNNNCCAACCGCrGrGrG||ACACTCTTTCCCTACACGACG/ideoxyU//ideoxyU/NNNNNNNNNN||CCAACCGC||rGrGrG | |||
|- | |||
| TSO.id14||B.TSO.N10.BC8.Idx14||ACACTCTTTCCCTACACGACG/ideoxyU//ideoxyU/NNNNNNNNNNGGATTACCrGrGrG||ACACTCTTTCCCTACACGACG/ideoxyU//ideoxyU/NNNNNNNNNN||GGATTACC||rGrGrG | |||
|- | |||
| TSO.id15||B.TSO.N10.BC8.Idx15||ACACTCTTTCCCTACACGACG/ideoxyU//ideoxyU/NNNNNNNNNNCAGAGACArGrGrG||ACACTCTTTCCCTACACGACG/ideoxyU//ideoxyU/NNNNNNNNNN||CAGAGACA||rGrGrG | |||
|- | |||
| TSO.id16||B.TSO.N10.BC8.Idx16||ACACTCTTTCCCTACACGACG/ideoxyU//ideoxyU/NNNNNNNNNNCAATAGTGrGrGrG||ACACTCTTTCCCTACACGACG/ideoxyU//ideoxyU/NNNNNNNNNN||CAATAGTG||rGrGrG | |||
|- | |||
| TSO.id17||B.TSO.N10.BC8.Idx17||ACACTCTTTCCCTACACGACG/ideoxyU//ideoxyU/NNNNNNNNNNGGTGGCCArGrGrG||ACACTCTTTCCCTACACGACG/ideoxyU//ideoxyU/NNNNNNNNNN||GGTGGCCA||rGrGrG | |||
|- | |||
| TSO.id18||B.TSO.N10.BC8.Idx18||ACACTCTTTCCCTACACGACG/ideoxyU//ideoxyU/NNNNNNNNNNGGAGGGAGrGrGrG||ACACTCTTTCCCTACACGACG/ideoxyU//ideoxyU/NNNNNNNNNN||GGAGGGAG||rGrGrG | |||
|- | |||
| TSO.id19||B.TSO.N10.BC8.Idx19||ACACTCTTTCCCTACACGACG/ideoxyU//ideoxyU/NNNNNNNNNNGTTGTCCTrGrGrG||ACACTCTTTCCCTACACGACG/ideoxyU//ideoxyU/NNNNNNNNNN||GTTGTCCT||rGrGrG | |||
|- | |||
| TSO.id20||B.TSO.N10.BC8.Idx20||ACACTCTTTCCCTACACGACG/ideoxyU//ideoxyU/NNNNNNNNNNTTCGACCTrGrGrG||ACACTCTTTCCCTACACGACG/ideoxyU//ideoxyU/NNNNNNNNNN||TTCGACCT||rGrGrG | |||
|- | |||
| TSO.id21||B.TSO.N10.BC8.Idx21||ACACTCTTTCCCTACACGACG/ideoxyU//ideoxyU/NNNNNNNNNNGCTTATTGrGrGrG||ACACTCTTTCCCTACACGACG/ideoxyU//ideoxyU/NNNNNNNNNN||GCTTATTG||rGrGrG | |||
|- | |||
| TSO.id22||B.TSO.N10.BC8.Idx22||ACACTCTTTCCCTACACGACG/ideoxyU//ideoxyU/NNNNNNNNNNCAACCTTTrGrGrG||ACACTCTTTCCCTACACGACG/ideoxyU//ideoxyU/NNNNNNNNNN||CAACCTTT||rGrGrG | |||
|- | |||
| TSO.id23||B.TSO.N10.BC8.Idx23||ACACTCTTTCCCTACACGACG/ideoxyU//ideoxyU/NNNNNNNNNNCTTAGGTArGrGrG||ACACTCTTTCCCTACACGACG/ideoxyU//ideoxyU/NNNNNNNNNN||CTTAGGTA||rGrGrG | |||
|- | |||
| TSO.id24||B.TSO.N10.BC8.Idx24||ACACTCTTTCCCTACACGACG/ideoxyU//ideoxyU/NNNNNNNNNNGCTCACCArGrGrG||ACACTCTTTCCCTACACGACG/ideoxyU//ideoxyU/NNNNNNNNNN||GCTCACCA||rGrGrG | |||
|- | |||
| TSO.id25||B.TSO.N10.BC8.Idx25||ACACTCTTTCCCTACACGACG/ideoxyU//ideoxyU/NNNNNNNNNNAGAAATGCrGrGrG||ACACTCTTTCCCTACACGACG/ideoxyU//ideoxyU/NNNNNNNNNN||AGAAATGC||rGrGrG | |||
|- | |||
| TSO.id26||B.TSO.N10.BC8.Idx26||ACACTCTTTCCCTACACGACG/ideoxyU//ideoxyU/NNNNNNNNNNCTTACTGArGrGrG||ACACTCTTTCCCTACACGACG/ideoxyU//ideoxyU/NNNNNNNNNN||CTTACTGA||rGrGrG | |||
|- | |||
| TSO.id27||B.TSO.N10.BC8.Idx27||ACACTCTTTCCCTACACGACG/ideoxyU//ideoxyU/NNNNNNNNNNCTCACGGGrGrGrG||ACACTCTTTCCCTACACGACG/ideoxyU//ideoxyU/NNNNNNNNNN||CTCACGGG||rGrGrG | |||
|- | |||
| TSO.id28||B.TSO.N10.BC8.Idx28||ACACTCTTTCCCTACACGACG/ideoxyU//ideoxyU/NNNNNNNNNNCACTGTCArGrGrG||ACACTCTTTCCCTACACGACG/ideoxyU//ideoxyU/NNNNNNNNNN||CACTGTCA||rGrGrG | |||
|- | |||
| TSO.id29||B.TSO.N10.BC8.Idx29||ACACTCTTTCCCTACACGACG/ideoxyU//ideoxyU/NNNNNNNNNNTTCTTGGGrGrGrG||ACACTCTTTCCCTACACGACG/ideoxyU//ideoxyU/NNNNNNNNNN||TTCTTGGG||rGrGrG | |||
|- | |||
| TSO.id30||B.TSO.N10.BC8.Idx30||ACACTCTTTCCCTACACGACG/ideoxyU//ideoxyU/NNNNNNNNNNACCGGCTTrGrGrG||ACACTCTTTCCCTACACGACG/ideoxyU//ideoxyU/NNNNNNNNNN||ACCGGCTT||rGrGrG | |||
|- | |||
| TSO.id31||B.TSO.N10.BC8.Idx31||ACACTCTTTCCCTACACGACG/ideoxyU//ideoxyU/NNNNNNNNNNTAACCACArGrGrG||ACACTCTTTCCCTACACGACG/ideoxyU//ideoxyU/NNNNNNNNNN||TAACCACA||rGrGrG | |||
|- | |||
| TSO.id32||B.TSO.N10.BC8.Idx32||ACACTCTTTCCCTACACGACG/ideoxyU//ideoxyU/NNNNNNNNNNTACGCGCGrGrGrG||ACACTCTTTCCCTACACGACG/ideoxyU//ideoxyU/NNNNNNNNNN||TACGCGCG||rGrGrG | |||
|- | |||
| TSO.id33||B.TSO.N10.BC8.Idx33||ACACTCTTTCCCTACACGACG/ideoxyU//ideoxyU/NNNNNNNNNNCCTCGATGrGrGrG||ACACTCTTTCCCTACACGACG/ideoxyU//ideoxyU/NNNNNNNNNN||CCTCGATG||rGrGrG | |||
|- | |||
| TSO.id34||B.TSO.N10.BC8.Idx34||ACACTCTTTCCCTACACGACG/ideoxyU//ideoxyU/NNNNNNNNNNAGAGCTCArGrGrG||ACACTCTTTCCCTACACGACG/ideoxyU//ideoxyU/NNNNNNNNNN||AGAGCTCA||rGrGrG | |||
|- | |||
| TSO.id35||B.TSO.N10.BC8.Idx35||ACACTCTTTCCCTACACGACG/ideoxyU//ideoxyU/NNNNNNNNNNTATGGTCCrGrGrG||ACACTCTTTCCCTACACGACG/ideoxyU//ideoxyU/NNNNNNNNNN||TATGGTCC||rGrGrG | |||
|- | |||
| TSO.id36||B.TSO.N10.BC8.Idx36||ACACTCTTTCCCTACACGACG/ideoxyU//ideoxyU/NNNNNNNNNNTGTCCGGCrGrGrG||ACACTCTTTCCCTACACGACG/ideoxyU//ideoxyU/NNNNNNNNNN||TGTCCGGC||rGrGrG | |||
|- | |||
| TSO.id37||B.TSO.N10.BC8.Idx37||ACACTCTTTCCCTACACGACG/ideoxyU//ideoxyU/NNNNNNNNNNAAGGAATCrGrGrG||ACACTCTTTCCCTACACGACG/ideoxyU//ideoxyU/NNNNNNNNNN||AAGGAATC||rGrGrG | |||
|- | |||
| TSO.id38||B.TSO.N10.BC8.Idx38||ACACTCTTTCCCTACACGACG/ideoxyU//ideoxyU/NNNNNNNNNNCATCGTAGrGrGrG||ACACTCTTTCCCTACACGACG/ideoxyU//ideoxyU/NNNNNNNNNN||CATCGTAG||rGrGrG | |||
|- | |||
| TSO.id39||B.TSO.N10.BC8.Idx39||ACACTCTTTCCCTACACGACG/ideoxyU//ideoxyU/NNNNNNNNNNGTGTTGAGrGrGrG||ACACTCTTTCCCTACACGACG/ideoxyU//ideoxyU/NNNNNNNNNN||GTGTTGAG||rGrGrG | |||
|- | |||
| TSO.id40||B.TSO.N10.BC8.Idx40||ACACTCTTTCCCTACACGACG/ideoxyU//ideoxyU/NNNNNNNNNNATGTATCCrGrGrG||ACACTCTTTCCCTACACGACG/ideoxyU//ideoxyU/NNNNNNNNNN||ATGTATCC||rGrGrG | |||
|- | |||
| TSO.id41||B.TSO.N10.BC8.Idx41||ACACTCTTTCCCTACACGACG/ideoxyU//ideoxyU/NNNNNNNNNNTGACAGGTrGrGrG||ACACTCTTTCCCTACACGACG/ideoxyU//ideoxyU/NNNNNNNNNN||TGACAGGT||rGrGrG | |||
|- | |||
| TSO.id42||B.TSO.N10.BC8.Idx42||ACACTCTTTCCCTACACGACG/ideoxyU//ideoxyU/NNNNNNNNNNTACGCCAArGrGrG||ACACTCTTTCCCTACACGACG/ideoxyU//ideoxyU/NNNNNNNNNN||TACGCCAA||rGrGrG | |||
|- | |||
| TSO.id43||B.TSO.N10.BC8.Idx43||ACACTCTTTCCCTACACGACG/ideoxyU//ideoxyU/NNNNNNNNNNGAAGGCAArGrGrG||ACACTCTTTCCCTACACGACG/ideoxyU//ideoxyU/NNNNNNNNNN||GAAGGCAA||rGrGrG | |||
|- | |||
| TSO.id44||B.TSO.N10.BC8.Idx44||ACACTCTTTCCCTACACGACG/ideoxyU//ideoxyU/NNNNNNNNNNCACCCGAGrGrGrG||ACACTCTTTCCCTACACGACG/ideoxyU//ideoxyU/NNNNNNNNNN||CACCCGAG||rGrGrG | |||
|- | |||
| TSO.id45||B.TSO.N10.BC8.Idx45||ACACTCTTTCCCTACACGACG/ideoxyU//ideoxyU/NNNNNNNNNNGGTGTGTCrGrGrG||ACACTCTTTCCCTACACGACG/ideoxyU//ideoxyU/NNNNNNNNNN||GGTGTGTC||rGrGrG | |||
|- | |||
| TSO.id46||B.TSO.N10.BC8.Idx46||ACACTCTTTCCCTACACGACG/ideoxyU//ideoxyU/NNNNNNNNNNTCTTGTGCrGrGrG||ACACTCTTTCCCTACACGACG/ideoxyU//ideoxyU/NNNNNNNNNN||TCTTGTGC||rGrGrG | |||
|- | |||
| TSO.id47||B.TSO.N10.BC8.Idx47||ACACTCTTTCCCTACACGACG/ideoxyU//ideoxyU/NNNNNNNNNNAGTACCTGrGrGrG||ACACTCTTTCCCTACACGACG/ideoxyU//ideoxyU/NNNNNNNNNN||AGTACCTG||rGrGrG | |||
|- | |||
| TSO.id48||B.TSO.N10.BC8.Idx48||ACACTCTTTCCCTACACGACG/ideoxyU//ideoxyU/NNNNNNNNNNAGCGTCCTrGrGrG||ACACTCTTTCCCTACACGACG/ideoxyU//ideoxyU/NNNNNNNNNN||AGCGTCCT||rGrGrG | |||
|- | |||
| PB_PCR.F||PB_PCR.F||AATGATACGGCGACCACCGAGATCTACACTCTTTCCCTACACGACG|||||| | |||
|} | |||
* The 3' adaptor list | |||
{| {{table}} border=1 | |||
| align="center" style="background:#f0f0f0;"|'''Tube ID''' | |||
| align="center" style="background:#f0f0f0;"|'''Primer full name''' | |||
| align="center" style="background:#f0f0f0;"|'''Primer full sequence''' | |||
| align="center" style="background:#f0f0f0;"|'''PE_R''' | |||
| align="center" style="background:#f0f0f0;"|'''Barcode''' | |||
| align="center" style="background:#f0f0f0;"|'''T20VN''' | |||
|- | |||
| T20.r01||T20VN_PE_R||/5Biosg/GCATTCCTGCTGAACCGCTCTTCCGATCTTTTTTTTTTTTTTTTTTTTTVN||/5Biosg/GCATTCCTGCTGAACCGCTCTTCCGATCT||\||TTTTTTTTTTTTTTTTTTTTVN | |||
|- | |||
| T20.r02||N9_PE_R||/5Biosg/GCATTCCTGCTGAACCGCTCTTCCGATCTNNNNNNNNN||/5Biosg/GCATTCCTGCTGAACCGCTCTTCCGATCT||\||NNNNNNNNN | |||
|- | |||
| T20.r03||T20VN_BC6_PE_R||GCATTCCTGCTGAACCGCTCTTTTTAGGTTTTTTTTTTTTTTTTTTTTTVN||/5Biosg/GCATTCCTGCTGAACCGCTCTT||TTTAGG||TTTTTTTTTTTTTTTTTTTTVN | |||
|- | |||
| T20.id01||T20VN.BC8.PE_R.Indx1||/5Biosg/GCATTCCTGCTGAACCGCTCTTCCGATCTTCTGAGACTTTTTTTTTTTTTTTTTTTTVN||/5Biosg/GCATTCCTGCTGAACCGCTCTTCCGATCT||TCTGAGAC||TTTTTTTTTTTTTTTTTTTTVN | |||
|- | |||
| T20.id02||T20VN.BC8.PE_R.Indx2||/5Biosg/GCATTCCTGCTGAACCGCTCTTCCGATCTTGCCCAAGTTTTTTTTTTTTTTTTTTTTVN||/5Biosg/GCATTCCTGCTGAACCGCTCTTCCGATCT||TGCCCAAG||TTTTTTTTTTTTTTTTTTTTVN | |||
|- | |||
| T20.id03||T20VN.BC8.PE_R.Indx3||/5Biosg/GCATTCCTGCTGAACCGCTCTTCCGATCTGACTCGCTTTTTTTTTTTTTTTTTTTTTVN||/5Biosg/GCATTCCTGCTGAACCGCTCTTCCGATCT||GACTCGCT||TTTTTTTTTTTTTTTTTTTTVN | |||
|- | |||
| T20.id04||T20VN.BC8.PE_R.Indx4||/5Biosg/GCATTCCTGCTGAACCGCTCTTCCGATCTCCACTCCATTTTTTTTTTTTTTTTTTTTVN||/5Biosg/GCATTCCTGCTGAACCGCTCTTCCGATCT||CCACTCCA||TTTTTTTTTTTTTTTTTTTTVN | |||
|- | |||
| T20.id05||T20VN.BC8.PE_R.Indx5||/5Biosg/GCATTCCTGCTGAACCGCTCTTCCGATCTGAACAGGGTTTTTTTTTTTTTTTTTTTTVN||/5Biosg/GCATTCCTGCTGAACCGCTCTTCCGATCT||GAACAGGG||TTTTTTTTTTTTTTTTTTTTVN | |||
|- | |||
| T20.id06||T20VN.BC8.PE_R.Indx6||/5Biosg/GCATTCCTGCTGAACCGCTCTTCCGATCTCTGCTGATTTTTTTTTTTTTTTTTTTTTVN||/5Biosg/GCATTCCTGCTGAACCGCTCTTCCGATCT||CTGCTGAT||TTTTTTTTTTTTTTTTTTTTVN | |||
|- | |||
| T20.id07||T20VN.BC8.PE_R.Indx7||/5Biosg/GCATTCCTGCTGAACCGCTCTTCCGATCTCTAGTATCTTTTTTTTTTTTTTTTTTTTVN||/5Biosg/GCATTCCTGCTGAACCGCTCTTCCGATCT||CTAGTATC||TTTTTTTTTTTTTTTTTTTTVN | |||
|- | |||
| T20.id08||T20VN.BC8.PE_R.Indx8||/5Biosg/GCATTCCTGCTGAACCGCTCTTCCGATCTGGCTGAAATTTTTTTTTTTTTTTTTTTTVN||/5Biosg/GCATTCCTGCTGAACCGCTCTTCCGATCT||GGCTGAAA||TTTTTTTTTTTTTTTTTTTTVN | |||
|- | |||
| T20.id09||T20VN.BC8.PE_R.Indx9||/5Biosg/GCATTCCTGCTGAACCGCTCTTCCGATCTGAGTGCTTTTTTTTTTTTTTTTTTTTTTVN||/5Biosg/GCATTCCTGCTGAACCGCTCTTCCGATCT||GAGTGCTT||TTTTTTTTTTTTTTTTTTTTVN | |||
|- | |||
| T20.id10||T20VN.BC8.PE_R.Indx10||/5Biosg/GCATTCCTGCTGAACCGCTCTTCCGATCTATACGCCATTTTTTTTTTTTTTTTTTTTVN||/5Biosg/GCATTCCTGCTGAACCGCTCTTCCGATCT||ATACGCCA||TTTTTTTTTTTTTTTTTTTTVN | |||
|- | |||
| T20.id11||T20VN.BC8.PE_R.Indx11||/5Biosg/GCATTCCTGCTGAACCGCTCTTCCGATCTATGCGTTATTTTTTTTTTTTTTTTTTTTVN||/5Biosg/GCATTCCTGCTGAACCGCTCTTCCGATCT||ATGCGTTA||TTTTTTTTTTTTTTTTTTTTVN | |||
|- | |||
| T20.id12||T20VN.BC8.PE_R.Indx12||/5Biosg/GCATTCCTGCTGAACCGCTCTTCCGATCTTCACCCTTTTTTTTTTTTTTTTTTTTTTVN||/5Biosg/GCATTCCTGCTGAACCGCTCTTCCGATCT||TCACCCTT||TTTTTTTTTTTTTTTTTTTTVN | |||
|- | |||
| T20.id13||T20V.BC8.PE_R.Indx13||/5Biosg/GCATTCCTGCTGAACCGCTCTTCCGATCTCCAACCGCTTTTTTTTTTTTTTTTTTTTV||/5Biosg/GCATTCCTGCTGAACCGCTCTTCCGATCT||CCAACCGC||TTTTTTTTTTTTTTTTTTTTV | |||
|- | |||
| T20.id14||T20V.BC8.PE_R.Indx14||/5Biosg/GCATTCCTGCTGAACCGCTCTTCCGATCTGGATTACCTTTTTTTTTTTTTTTTTTTTV||/5Biosg/GCATTCCTGCTGAACCGCTCTTCCGATCT||GGATTACC||TTTTTTTTTTTTTTTTTTTTV | |||
|- | |||
| T20.id15||T20V.BC8.PE_R.Indx15||/5Biosg/GCATTCCTGCTGAACCGCTCTTCCGATCTCAGAGACATTTTTTTTTTTTTTTTTTTTV||/5Biosg/GCATTCCTGCTGAACCGCTCTTCCGATCT||CAGAGACA||TTTTTTTTTTTTTTTTTTTTV | |||
|- | |||
| T20.id16||T20V.BC8.PE_R.Indx16||/5Biosg/GCATTCCTGCTGAACCGCTCTTCCGATCTCAATAGTGTTTTTTTTTTTTTTTTTTTTV||/5Biosg/GCATTCCTGCTGAACCGCTCTTCCGATCT||CAATAGTG||TTTTTTTTTTTTTTTTTTTTV | |||
|- | |||
| T20.id17||T20V.BC8.PE_R.Indx17||/5Biosg/GCATTCCTGCTGAACCGCTCTTCCGATCTGGTGGCCATTTTTTTTTTTTTTTTTTTTV||/5Biosg/GCATTCCTGCTGAACCGCTCTTCCGATCT||GGTGGCCA||TTTTTTTTTTTTTTTTTTTTV | |||
|- | |||
| T20.id18||T20V.BC8.PE_R.Indx18||/5Biosg/GCATTCCTGCTGAACCGCTCTTCCGATCTGGAGGGAGTTTTTTTTTTTTTTTTTTTTV||/5Biosg/GCATTCCTGCTGAACCGCTCTTCCGATCT||GGAGGGAG||TTTTTTTTTTTTTTTTTTTTV | |||
|- | |||
| T20.id19||T20V.BC8.PE_R.Indx19||/5Biosg/GCATTCCTGCTGAACCGCTCTTCCGATCTGTTGTCCTTTTTTTTTTTTTTTTTTTTTV||/5Biosg/GCATTCCTGCTGAACCGCTCTTCCGATCT||GTTGTCCT||TTTTTTTTTTTTTTTTTTTTV | |||
|- | |||
| T20.id20||T20V.BC8.PE_R.Indx20||/5Biosg/GCATTCCTGCTGAACCGCTCTTCCGATCTTTCGACCTTTTTTTTTTTTTTTTTTTTTV||/5Biosg/GCATTCCTGCTGAACCGCTCTTCCGATCT||TTCGACCT||TTTTTTTTTTTTTTTTTTTTV | |||
|- | |||
| T20.id21||T20V.BC8.PE_R.Indx21||/5Biosg/GCATTCCTGCTGAACCGCTCTTCCGATCTGCTTATTGTTTTTTTTTTTTTTTTTTTTV||/5Biosg/GCATTCCTGCTGAACCGCTCTTCCGATCT||GCTTATTG||TTTTTTTTTTTTTTTTTTTTV | |||
|- | |||
| T20.id22||T20V.BC8.PE_R.Indx22||/5Biosg/GCATTCCTGCTGAACCGCTCTTCCGATCTCAACCTTTTTTTTTTTTTTTTTTTTTTTV||/5Biosg/GCATTCCTGCTGAACCGCTCTTCCGATCT||CAACCTTT||TTTTTTTTTTTTTTTTTTTTV | |||
|- | |||
| T20.id23||T20V.BC8.PE_R.Indx23||/5Biosg/GCATTCCTGCTGAACCGCTCTTCCGATCTCTTAGGTATTTTTTTTTTTTTTTTTTTTV||/5Biosg/GCATTCCTGCTGAACCGCTCTTCCGATCT||CTTAGGTA||TTTTTTTTTTTTTTTTTTTTV | |||
|- | |||
| T20.id24||T20V.BC8.PE_R.Indx24||/5Biosg/GCATTCCTGCTGAACCGCTCTTCCGATCTGCTCACCATTTTTTTTTTTTTTTTTTTTV||/5Biosg/GCATTCCTGCTGAACCGCTCTTCCGATCT||GCTCACCA||TTTTTTTTTTTTTTTTTTTTV | |||
|- | |||
| N2.id73||PCR_R.N2Ind73||CAAGCAGAAGACGGCATACGAGATGAGTCCCTCGGCATTCCTGCTGAACCGCTCTT||CAAGCAGAAGACGGCATACGAGAT||GAGTCC||CTCGGCATTCCTGCTGAACCGCTCTT | |||
|- | |||
| N2.id74||PCR_R.N2Ind74||CAAGCAGAAGACGGCATACGAGATTCGCCACTCGGCATTCCTGCTGAACCGCTCTT||CAAGCAGAAGACGGCATACGAGAT||TCGCCA||CTCGGCATTCCTGCTGAACCGCTCTT | |||
|- | |||
| N2.id75||PCR_R.N2Ind75||CAAGCAGAAGACGGCATACGAGATAAGTCGCTCGGCATTCCTGCTGAACCGCTCTT||CAAGCAGAAGACGGCATACGAGAT||AAGTCG||CTCGGCATTCCTGCTGAACCGCTCTT | |||
|- | |||
| N2.id76||PCR_R.N2Ind76||CAAGCAGAAGACGGCATACGAGATAATAGGCTCGGCATTCCTGCTGAACCGCTCTT||CAAGCAGAAGACGGCATACGAGAT||AATAGG||CTCGGCATTCCTGCTGAACCGCTCTT | |||
|- | |||
| N2.id77||PCR_R.N2Ind77||CAAGCAGAAGACGGCATACGAGATACCCGTCTCGGCATTCCTGCTGAACCGCTCTT||CAAGCAGAAGACGGCATACGAGAT||ACCCGT||CTCGGCATTCCTGCTGAACCGCTCTT | |||
|- | |||
| N2.id78||PCR_R.N2Ind78||CAAGCAGAAGACGGCATACGAGATAACACGCTCGGCATTCCTGCTGAACCGCTCTT||CAAGCAGAAGACGGCATACGAGAT||AACACG||CTCGGCATTCCTGCTGAACCGCTCTT | |||
|- | |||
| N2.id79||PCR_R.N2Ind79||CAAGCAGAAGACGGCATACGAGATGCTTGGCTCGGCATTCCTGCTGAACCGCTCTT||CAAGCAGAAGACGGCATACGAGAT||GCTTGG||CTCGGCATTCCTGCTGAACCGCTCTT | |||
|- | |||
| N2.id80||PCR_R.N2Ind80||CAAGCAGAAGACGGCATACGAGATTTACCACTCGGCATTCCTGCTGAACCGCTCTT||CAAGCAGAAGACGGCATACGAGAT||TTACCA||CTCGGCATTCCTGCTGAACCGCTCTT | |||
|- | |||
| N2.id81||PCR_R.N2Ind81||CAAGCAGAAGACGGCATACGAGATCCAGGTCTCGGCATTCCTGCTGAACCGCTCTT||CAAGCAGAAGACGGCATACGAGAT||CCAGGT||CTCGGCATTCCTGCTGAACCGCTCTT | |||
|- | |||
| N2.id82||PCR_R.N2Ind82||CAAGCAGAAGACGGCATACGAGATCGTTTGCTCGGCATTCCTGCTGAACCGCTCTT||CAAGCAGAAGACGGCATACGAGAT||CGTTTG||CTCGGCATTCCTGCTGAACCGCTCTT | |||
|- | |||
| N2.id83||PCR_R.N2Ind83||CAAGCAGAAGACGGCATACGAGATGACCACCTCGGCATTCCTGCTGAACCGCTCTT||CAAGCAGAAGACGGCATACGAGAT||GACCAC||CTCGGCATTCCTGCTGAACCGCTCTT | |||
|- | |||
| N2.id84||PCR_R.N2Ind84||CAAGCAGAAGACGGCATACGAGATACAAGACTCGGCATTCCTGCTGAACCGCTCTT||CAAGCAGAAGACGGCATACGAGAT||ACAAGA||CTCGGCATTCCTGCTGAACCGCTCTT | |||
|- | |||
| N2.id85||PCR_R.N2Ind85||CAAGCAGAAGACGGCATACGAGATACCGCACTCGGCATTCCTGCTGAACCGCTCTT||CAAGCAGAAGACGGCATACGAGAT||ACCGCA||CTCGGCATTCCTGCTGAACCGCTCTT | |||
|- | |||
| N2.id86||PCR_R.N2Ind86||CAAGCAGAAGACGGCATACGAGATTGGGTACTCGGCATTCCTGCTGAACCGCTCTT||CAAGCAGAAGACGGCATACGAGAT||TGGGTA||CTCGGCATTCCTGCTGAACCGCTCTT | |||
|- | |||
| N2.id87||PCR_R.N2Ind87||CAAGCAGAAGACGGCATACGAGATATTCCGCTCGGCATTCCTGCTGAACCGCTCTT||CAAGCAGAAGACGGCATACGAGAT||ATTCCG||CTCGGCATTCCTGCTGAACCGCTCTT | |||
|- | |||
| N2.id88||PCR_R.N2Ind88||CAAGCAGAAGACGGCATACGAGATGAATGTCTCGGCATTCCTGCTGAACCGCTCTT||CAAGCAGAAGACGGCATACGAGAT||GAATGT||CTCGGCATTCCTGCTGAACCGCTCTT | |||
|- | |||
| N2.id89||PCR_R.N2Ind89||CAAGCAGAAGACGGCATACGAGATGCTGATCTCGGCATTCCTGCTGAACCGCTCTT||CAAGCAGAAGACGGCATACGAGAT||GCTGAT||CTCGGCATTCCTGCTGAACCGCTCTT | |||
|- | |||
| N2.id90||PCR_R.N2Ind90||CAAGCAGAAGACGGCATACGAGATAGTGCTCTCGGCATTCCTGCTGAACCGCTCTT||CAAGCAGAAGACGGCATACGAGAT||AGTGCT||CTCGGCATTCCTGCTGAACCGCTCTT | |||
|- | |||
| N2.id91||PCR_R.N2Ind91||CAAGCAGAAGACGGCATACGAGATCAGGGACTCGGCATTCCTGCTGAACCGCTCTT||CAAGCAGAAGACGGCATACGAGAT||CAGGGA||CTCGGCATTCCTGCTGAACCGCTCTT | |||
|- | |||
| N2.id92||PCR_R.N2Ind92||CAAGCAGAAGACGGCATACGAGATCATGCGCTCGGCATTCCTGCTGAACCGCTCTT||CAAGCAGAAGACGGCATACGAGAT||CATGCG||CTCGGCATTCCTGCTGAACCGCTCTT | |||
|- | |||
| N2.id93||PCR_R.N2Ind93||CAAGCAGAAGACGGCATACGAGATTGCCTACTCGGCATTCCTGCTGAACCGCTCTT||CAAGCAGAAGACGGCATACGAGAT||TGCCTA||CTCGGCATTCCTGCTGAACCGCTCTT | |||
|- | |||
| N2.id94||PCR_R.N2Ind94||CAAGCAGAAGACGGCATACGAGATCTATACCTCGGCATTCCTGCTGAACCGCTCTT||CAAGCAGAAGACGGCATACGAGAT||CTATAC||CTCGGCATTCCTGCTGAACCGCTCTT | |||
|- | |||
| N2.id95||PCR_R.N2Ind95||CAAGCAGAAGACGGCATACGAGATCCGAGTCTCGGCATTCCTGCTGAACCGCTCTT||CAAGCAGAAGACGGCATACGAGAT||CCGAGT||CTCGGCATTCCTGCTGAACCGCTCTT | |||
|- | |||
| N2.id96||PCR_R.N2Ind96||CAAGCAGAAGACGGCATACGAGATACCTGCCTCGGCATTCCTGCTGAACCGCTCTT||CAAGCAGAAGACGGCATACGAGAT||ACCTGC||CTCGGCATTCCTGCTGAACCGCTCTT | |||
|} | |||
==Sequencing primers== | |||
totoRNAseq Read 1: [http://genome-tech.ucsd.edu/LabNotes/index.php/Rui:LabNotes/SingleCell/2012-10-30#Sequencing_primer_design] | |||
CACCGAGATCTACACTCTTTCCCTACACGACGTT | |||
34nt Tm=64(IDT) Tm=71.72(Kun's) | |||
correspond to TSO.idXX primers used for 5'end | |||
totoRNAseq Read 1 v2: | |||
CCACCGAGATCTACACTCTTTCCCTACACGACG | |||
33nt Tm=64.5(IDT) Tm=72.04(Kun's) | |||
correspond to TSO.r02/TSO.r04 primers used for 5'end |
Latest revision as of 21:21, 6 March 2013
totoRNA library structure[edit]
Standard primers[edit]
totoRNAseq primers[edit]
File:Libraries totoRNA-5.jpg File:Libraries totoRNA-3.jpg
totoRNA primer sequence[edit]
- The 5' adaptors list
Tube ID | Primer full name | Primer full sequence | PE_F-N10 | Barcode | TSO |
TSO.r01 | BR_TSO_N10_BC6 | ACACTCTTTCCCTACACGACGCTCTTCCGATCTNNNdUNNNNdUNNNTTTAGGrGrGrG | ACACTCTTTCCCTACACGACGCTCTTCCGATCTNNN/ideoxyU/NNNN/ideoxyU/NNN | TTTAGG | rGrGrG |
TSO.r02 | B_TSO_N10 | ACACTCTTTCCCTACACGACGNNNUNNNNUNNNrGrGrG | ACACTCTTTCCCTACACGACGNNN/ideoxyU/NNNN/ideoxyU/NNN | \ | rGrGrG |
TSO.r03 | B.N10.TSO.NH2 | ACACTCTTTCCCTACACGACGUNNNUNNNNUNNNrGrGrG/3AmMO/ | ACACTCTTTCCCTACACGACGNNN/ideoxyU/NNNN/ideoxyU/NNN | \ | rGrGrG |
TSO.id01 | B.TSO.N10.BC8.Idx1 | ACACTCTTTCCCTACACGACG/ideoxyU//ideoxyU/NNNNNNNNNNTCTGAGACrGrGrG | ACACTCTTTCCCTACACGACG/ideoxyU//ideoxyU/NNNNNNNNNN | TCTGAGAC | rGrGrG |
TSO.id02 | B.TSO.N10.BC8.Idx2 | ACACTCTTTCCCTACACGACG/ideoxyU//ideoxyU/NNNNNNNNNNTGCCCAAGrGrGrG | ACACTCTTTCCCTACACGACG/ideoxyU//ideoxyU/NNNNNNNNNN | TGCCCAAG | rGrGrG |
TSO.id03 | B.TSO.N10.BC8.Idx3 | ACACTCTTTCCCTACACGACG/ideoxyU//ideoxyU/NNNNNNNNNNGACTCGCTrGrGrG | ACACTCTTTCCCTACACGACG/ideoxyU//ideoxyU/NNNNNNNNNN | GACTCGCT | rGrGrG |
TSO.id04 | B.TSO.N10.BC8.Idx4 | ACACTCTTTCCCTACACGACG/ideoxyU//ideoxyU/NNNNNNNNNNCCACTCCArGrGrG | ACACTCTTTCCCTACACGACG/ideoxyU//ideoxyU/NNNNNNNNNN | CCACTCCA | rGrGrG |
TSO.id05 | B.TSO.N10.BC8.Idx5 | ACACTCTTTCCCTACACGACG/ideoxyU//ideoxyU/NNNNNNNNNNGAACAGGGrGrGrG | ACACTCTTTCCCTACACGACG/ideoxyU//ideoxyU/NNNNNNNNNN | GAACAGGG | rGrGrG |
TSO.id06 | B.TSO.N10.BC8.Idx6 | ACACTCTTTCCCTACACGACG/ideoxyU//ideoxyU/NNNNNNNNNNCTGCTGATrGrGrG | ACACTCTTTCCCTACACGACG/ideoxyU//ideoxyU/NNNNNNNNNN | CTGCTGAT | rGrGrG |
TSO.id07 | B.TSO.N10.BC8.Idx7 | ACACTCTTTCCCTACACGACG/ideoxyU//ideoxyU/NNNNNNNNNNCTAGTATCrGrGrG | ACACTCTTTCCCTACACGACG/ideoxyU//ideoxyU/NNNNNNNNNN | CTAGTATC | rGrGrG |
TSO.id08 | B.TSO.N10.BC8.Idx8 | ACACTCTTTCCCTACACGACG/ideoxyU//ideoxyU/NNNNNNNNNNGGCTGAAArGrGrG | ACACTCTTTCCCTACACGACG/ideoxyU//ideoxyU/NNNNNNNNNN | GGCTGAAA | rGrGrG |
TSO.id09 | B.TSO.N10.BC8.Idx9 | ACACTCTTTCCCTACACGACG/ideoxyU//ideoxyU/NNNNNNNNNNGAGTGCTTrGrGrG | ACACTCTTTCCCTACACGACG/ideoxyU//ideoxyU/NNNNNNNNNN | GAGTGCTT | rGrGrG |
TSO.id10 | B.TSO.N10.BC8.Idx10 | ACACTCTTTCCCTACACGACG/ideoxyU//ideoxyU/NNNNNNNNNNATACGCCArGrGrG | ACACTCTTTCCCTACACGACG/ideoxyU//ideoxyU/NNNNNNNNNN | ATACGCCA | rGrGrG |
TSO.id11 | B.TSO.N10.BC8.Idx11 | ACACTCTTTCCCTACACGACG/ideoxyU//ideoxyU/NNNNNNNNNNATGCGTTArGrGrG | ACACTCTTTCCCTACACGACG/ideoxyU//ideoxyU/NNNNNNNNNN | ATGCGTTA | rGrGrG |
TSO.id12 | B.TSO.N10.BC8.Idx12 | ACACTCTTTCCCTACACGACG/ideoxyU//ideoxyU/NNNNNNNNNNTCACCCTTrGrGrG | ACACTCTTTCCCTACACGACG/ideoxyU//ideoxyU/NNNNNNNNNN | TCACCCTT | rGrGrG |
TSO.id13 | B.TSO.N10.BC8.Idx13 | ACACTCTTTCCCTACACGACG/ideoxyU//ideoxyU/NNNNNNNNNNCCAACCGCrGrGrG | ACACTCTTTCCCTACACGACG/ideoxyU//ideoxyU/NNNNNNNNNN | CCAACCGC | rGrGrG |
TSO.id14 | B.TSO.N10.BC8.Idx14 | ACACTCTTTCCCTACACGACG/ideoxyU//ideoxyU/NNNNNNNNNNGGATTACCrGrGrG | ACACTCTTTCCCTACACGACG/ideoxyU//ideoxyU/NNNNNNNNNN | GGATTACC | rGrGrG |
TSO.id15 | B.TSO.N10.BC8.Idx15 | ACACTCTTTCCCTACACGACG/ideoxyU//ideoxyU/NNNNNNNNNNCAGAGACArGrGrG | ACACTCTTTCCCTACACGACG/ideoxyU//ideoxyU/NNNNNNNNNN | CAGAGACA | rGrGrG |
TSO.id16 | B.TSO.N10.BC8.Idx16 | ACACTCTTTCCCTACACGACG/ideoxyU//ideoxyU/NNNNNNNNNNCAATAGTGrGrGrG | ACACTCTTTCCCTACACGACG/ideoxyU//ideoxyU/NNNNNNNNNN | CAATAGTG | rGrGrG |
TSO.id17 | B.TSO.N10.BC8.Idx17 | ACACTCTTTCCCTACACGACG/ideoxyU//ideoxyU/NNNNNNNNNNGGTGGCCArGrGrG | ACACTCTTTCCCTACACGACG/ideoxyU//ideoxyU/NNNNNNNNNN | GGTGGCCA | rGrGrG |
TSO.id18 | B.TSO.N10.BC8.Idx18 | ACACTCTTTCCCTACACGACG/ideoxyU//ideoxyU/NNNNNNNNNNGGAGGGAGrGrGrG | ACACTCTTTCCCTACACGACG/ideoxyU//ideoxyU/NNNNNNNNNN | GGAGGGAG | rGrGrG |
TSO.id19 | B.TSO.N10.BC8.Idx19 | ACACTCTTTCCCTACACGACG/ideoxyU//ideoxyU/NNNNNNNNNNGTTGTCCTrGrGrG | ACACTCTTTCCCTACACGACG/ideoxyU//ideoxyU/NNNNNNNNNN | GTTGTCCT | rGrGrG |
TSO.id20 | B.TSO.N10.BC8.Idx20 | ACACTCTTTCCCTACACGACG/ideoxyU//ideoxyU/NNNNNNNNNNTTCGACCTrGrGrG | ACACTCTTTCCCTACACGACG/ideoxyU//ideoxyU/NNNNNNNNNN | TTCGACCT | rGrGrG |
TSO.id21 | B.TSO.N10.BC8.Idx21 | ACACTCTTTCCCTACACGACG/ideoxyU//ideoxyU/NNNNNNNNNNGCTTATTGrGrGrG | ACACTCTTTCCCTACACGACG/ideoxyU//ideoxyU/NNNNNNNNNN | GCTTATTG | rGrGrG |
TSO.id22 | B.TSO.N10.BC8.Idx22 | ACACTCTTTCCCTACACGACG/ideoxyU//ideoxyU/NNNNNNNNNNCAACCTTTrGrGrG | ACACTCTTTCCCTACACGACG/ideoxyU//ideoxyU/NNNNNNNNNN | CAACCTTT | rGrGrG |
TSO.id23 | B.TSO.N10.BC8.Idx23 | ACACTCTTTCCCTACACGACG/ideoxyU//ideoxyU/NNNNNNNNNNCTTAGGTArGrGrG | ACACTCTTTCCCTACACGACG/ideoxyU//ideoxyU/NNNNNNNNNN | CTTAGGTA | rGrGrG |
TSO.id24 | B.TSO.N10.BC8.Idx24 | ACACTCTTTCCCTACACGACG/ideoxyU//ideoxyU/NNNNNNNNNNGCTCACCArGrGrG | ACACTCTTTCCCTACACGACG/ideoxyU//ideoxyU/NNNNNNNNNN | GCTCACCA | rGrGrG |
TSO.id25 | B.TSO.N10.BC8.Idx25 | ACACTCTTTCCCTACACGACG/ideoxyU//ideoxyU/NNNNNNNNNNAGAAATGCrGrGrG | ACACTCTTTCCCTACACGACG/ideoxyU//ideoxyU/NNNNNNNNNN | AGAAATGC | rGrGrG |
TSO.id26 | B.TSO.N10.BC8.Idx26 | ACACTCTTTCCCTACACGACG/ideoxyU//ideoxyU/NNNNNNNNNNCTTACTGArGrGrG | ACACTCTTTCCCTACACGACG/ideoxyU//ideoxyU/NNNNNNNNNN | CTTACTGA | rGrGrG |
TSO.id27 | B.TSO.N10.BC8.Idx27 | ACACTCTTTCCCTACACGACG/ideoxyU//ideoxyU/NNNNNNNNNNCTCACGGGrGrGrG | ACACTCTTTCCCTACACGACG/ideoxyU//ideoxyU/NNNNNNNNNN | CTCACGGG | rGrGrG |
TSO.id28 | B.TSO.N10.BC8.Idx28 | ACACTCTTTCCCTACACGACG/ideoxyU//ideoxyU/NNNNNNNNNNCACTGTCArGrGrG | ACACTCTTTCCCTACACGACG/ideoxyU//ideoxyU/NNNNNNNNNN | CACTGTCA | rGrGrG |
TSO.id29 | B.TSO.N10.BC8.Idx29 | ACACTCTTTCCCTACACGACG/ideoxyU//ideoxyU/NNNNNNNNNNTTCTTGGGrGrGrG | ACACTCTTTCCCTACACGACG/ideoxyU//ideoxyU/NNNNNNNNNN | TTCTTGGG | rGrGrG |
TSO.id30 | B.TSO.N10.BC8.Idx30 | ACACTCTTTCCCTACACGACG/ideoxyU//ideoxyU/NNNNNNNNNNACCGGCTTrGrGrG | ACACTCTTTCCCTACACGACG/ideoxyU//ideoxyU/NNNNNNNNNN | ACCGGCTT | rGrGrG |
TSO.id31 | B.TSO.N10.BC8.Idx31 | ACACTCTTTCCCTACACGACG/ideoxyU//ideoxyU/NNNNNNNNNNTAACCACArGrGrG | ACACTCTTTCCCTACACGACG/ideoxyU//ideoxyU/NNNNNNNNNN | TAACCACA | rGrGrG |
TSO.id32 | B.TSO.N10.BC8.Idx32 | ACACTCTTTCCCTACACGACG/ideoxyU//ideoxyU/NNNNNNNNNNTACGCGCGrGrGrG | ACACTCTTTCCCTACACGACG/ideoxyU//ideoxyU/NNNNNNNNNN | TACGCGCG | rGrGrG |
TSO.id33 | B.TSO.N10.BC8.Idx33 | ACACTCTTTCCCTACACGACG/ideoxyU//ideoxyU/NNNNNNNNNNCCTCGATGrGrGrG | ACACTCTTTCCCTACACGACG/ideoxyU//ideoxyU/NNNNNNNNNN | CCTCGATG | rGrGrG |
TSO.id34 | B.TSO.N10.BC8.Idx34 | ACACTCTTTCCCTACACGACG/ideoxyU//ideoxyU/NNNNNNNNNNAGAGCTCArGrGrG | ACACTCTTTCCCTACACGACG/ideoxyU//ideoxyU/NNNNNNNNNN | AGAGCTCA | rGrGrG |
TSO.id35 | B.TSO.N10.BC8.Idx35 | ACACTCTTTCCCTACACGACG/ideoxyU//ideoxyU/NNNNNNNNNNTATGGTCCrGrGrG | ACACTCTTTCCCTACACGACG/ideoxyU//ideoxyU/NNNNNNNNNN | TATGGTCC | rGrGrG |
TSO.id36 | B.TSO.N10.BC8.Idx36 | ACACTCTTTCCCTACACGACG/ideoxyU//ideoxyU/NNNNNNNNNNTGTCCGGCrGrGrG | ACACTCTTTCCCTACACGACG/ideoxyU//ideoxyU/NNNNNNNNNN | TGTCCGGC | rGrGrG |
TSO.id37 | B.TSO.N10.BC8.Idx37 | ACACTCTTTCCCTACACGACG/ideoxyU//ideoxyU/NNNNNNNNNNAAGGAATCrGrGrG | ACACTCTTTCCCTACACGACG/ideoxyU//ideoxyU/NNNNNNNNNN | AAGGAATC | rGrGrG |
TSO.id38 | B.TSO.N10.BC8.Idx38 | ACACTCTTTCCCTACACGACG/ideoxyU//ideoxyU/NNNNNNNNNNCATCGTAGrGrGrG | ACACTCTTTCCCTACACGACG/ideoxyU//ideoxyU/NNNNNNNNNN | CATCGTAG | rGrGrG |
TSO.id39 | B.TSO.N10.BC8.Idx39 | ACACTCTTTCCCTACACGACG/ideoxyU//ideoxyU/NNNNNNNNNNGTGTTGAGrGrGrG | ACACTCTTTCCCTACACGACG/ideoxyU//ideoxyU/NNNNNNNNNN | GTGTTGAG | rGrGrG |
TSO.id40 | B.TSO.N10.BC8.Idx40 | ACACTCTTTCCCTACACGACG/ideoxyU//ideoxyU/NNNNNNNNNNATGTATCCrGrGrG | ACACTCTTTCCCTACACGACG/ideoxyU//ideoxyU/NNNNNNNNNN | ATGTATCC | rGrGrG |
TSO.id41 | B.TSO.N10.BC8.Idx41 | ACACTCTTTCCCTACACGACG/ideoxyU//ideoxyU/NNNNNNNNNNTGACAGGTrGrGrG | ACACTCTTTCCCTACACGACG/ideoxyU//ideoxyU/NNNNNNNNNN | TGACAGGT | rGrGrG |
TSO.id42 | B.TSO.N10.BC8.Idx42 | ACACTCTTTCCCTACACGACG/ideoxyU//ideoxyU/NNNNNNNNNNTACGCCAArGrGrG | ACACTCTTTCCCTACACGACG/ideoxyU//ideoxyU/NNNNNNNNNN | TACGCCAA | rGrGrG |
TSO.id43 | B.TSO.N10.BC8.Idx43 | ACACTCTTTCCCTACACGACG/ideoxyU//ideoxyU/NNNNNNNNNNGAAGGCAArGrGrG | ACACTCTTTCCCTACACGACG/ideoxyU//ideoxyU/NNNNNNNNNN | GAAGGCAA | rGrGrG |
TSO.id44 | B.TSO.N10.BC8.Idx44 | ACACTCTTTCCCTACACGACG/ideoxyU//ideoxyU/NNNNNNNNNNCACCCGAGrGrGrG | ACACTCTTTCCCTACACGACG/ideoxyU//ideoxyU/NNNNNNNNNN | CACCCGAG | rGrGrG |
TSO.id45 | B.TSO.N10.BC8.Idx45 | ACACTCTTTCCCTACACGACG/ideoxyU//ideoxyU/NNNNNNNNNNGGTGTGTCrGrGrG | ACACTCTTTCCCTACACGACG/ideoxyU//ideoxyU/NNNNNNNNNN | GGTGTGTC | rGrGrG |
TSO.id46 | B.TSO.N10.BC8.Idx46 | ACACTCTTTCCCTACACGACG/ideoxyU//ideoxyU/NNNNNNNNNNTCTTGTGCrGrGrG | ACACTCTTTCCCTACACGACG/ideoxyU//ideoxyU/NNNNNNNNNN | TCTTGTGC | rGrGrG |
TSO.id47 | B.TSO.N10.BC8.Idx47 | ACACTCTTTCCCTACACGACG/ideoxyU//ideoxyU/NNNNNNNNNNAGTACCTGrGrGrG | ACACTCTTTCCCTACACGACG/ideoxyU//ideoxyU/NNNNNNNNNN | AGTACCTG | rGrGrG |
TSO.id48 | B.TSO.N10.BC8.Idx48 | ACACTCTTTCCCTACACGACG/ideoxyU//ideoxyU/NNNNNNNNNNAGCGTCCTrGrGrG | ACACTCTTTCCCTACACGACG/ideoxyU//ideoxyU/NNNNNNNNNN | AGCGTCCT | rGrGrG |
PB_PCR.F | PB_PCR.F | AATGATACGGCGACCACCGAGATCTACACTCTTTCCCTACACGACG |
- The 3' adaptor list
Tube ID | Primer full name | Primer full sequence | PE_R | Barcode | T20VN |
T20.r01 | T20VN_PE_R | /5Biosg/GCATTCCTGCTGAACCGCTCTTCCGATCTTTTTTTTTTTTTTTTTTTTTVN | /5Biosg/GCATTCCTGCTGAACCGCTCTTCCGATCT | \ | TTTTTTTTTTTTTTTTTTTTVN |
T20.r02 | N9_PE_R | /5Biosg/GCATTCCTGCTGAACCGCTCTTCCGATCTNNNNNNNNN | /5Biosg/GCATTCCTGCTGAACCGCTCTTCCGATCT | \ | NNNNNNNNN |
T20.r03 | T20VN_BC6_PE_R | GCATTCCTGCTGAACCGCTCTTTTTAGGTTTTTTTTTTTTTTTTTTTTTVN | /5Biosg/GCATTCCTGCTGAACCGCTCTT | TTTAGG | TTTTTTTTTTTTTTTTTTTTVN |
T20.id01 | T20VN.BC8.PE_R.Indx1 | /5Biosg/GCATTCCTGCTGAACCGCTCTTCCGATCTTCTGAGACTTTTTTTTTTTTTTTTTTTTVN | /5Biosg/GCATTCCTGCTGAACCGCTCTTCCGATCT | TCTGAGAC | TTTTTTTTTTTTTTTTTTTTVN |
T20.id02 | T20VN.BC8.PE_R.Indx2 | /5Biosg/GCATTCCTGCTGAACCGCTCTTCCGATCTTGCCCAAGTTTTTTTTTTTTTTTTTTTTVN | /5Biosg/GCATTCCTGCTGAACCGCTCTTCCGATCT | TGCCCAAG | TTTTTTTTTTTTTTTTTTTTVN |
T20.id03 | T20VN.BC8.PE_R.Indx3 | /5Biosg/GCATTCCTGCTGAACCGCTCTTCCGATCTGACTCGCTTTTTTTTTTTTTTTTTTTTTVN | /5Biosg/GCATTCCTGCTGAACCGCTCTTCCGATCT | GACTCGCT | TTTTTTTTTTTTTTTTTTTTVN |
T20.id04 | T20VN.BC8.PE_R.Indx4 | /5Biosg/GCATTCCTGCTGAACCGCTCTTCCGATCTCCACTCCATTTTTTTTTTTTTTTTTTTTVN | /5Biosg/GCATTCCTGCTGAACCGCTCTTCCGATCT | CCACTCCA | TTTTTTTTTTTTTTTTTTTTVN |
T20.id05 | T20VN.BC8.PE_R.Indx5 | /5Biosg/GCATTCCTGCTGAACCGCTCTTCCGATCTGAACAGGGTTTTTTTTTTTTTTTTTTTTVN | /5Biosg/GCATTCCTGCTGAACCGCTCTTCCGATCT | GAACAGGG | TTTTTTTTTTTTTTTTTTTTVN |
T20.id06 | T20VN.BC8.PE_R.Indx6 | /5Biosg/GCATTCCTGCTGAACCGCTCTTCCGATCTCTGCTGATTTTTTTTTTTTTTTTTTTTTVN | /5Biosg/GCATTCCTGCTGAACCGCTCTTCCGATCT | CTGCTGAT | TTTTTTTTTTTTTTTTTTTTVN |
T20.id07 | T20VN.BC8.PE_R.Indx7 | /5Biosg/GCATTCCTGCTGAACCGCTCTTCCGATCTCTAGTATCTTTTTTTTTTTTTTTTTTTTVN | /5Biosg/GCATTCCTGCTGAACCGCTCTTCCGATCT | CTAGTATC | TTTTTTTTTTTTTTTTTTTTVN |
T20.id08 | T20VN.BC8.PE_R.Indx8 | /5Biosg/GCATTCCTGCTGAACCGCTCTTCCGATCTGGCTGAAATTTTTTTTTTTTTTTTTTTTVN | /5Biosg/GCATTCCTGCTGAACCGCTCTTCCGATCT | GGCTGAAA | TTTTTTTTTTTTTTTTTTTTVN |
T20.id09 | T20VN.BC8.PE_R.Indx9 | /5Biosg/GCATTCCTGCTGAACCGCTCTTCCGATCTGAGTGCTTTTTTTTTTTTTTTTTTTTTTVN | /5Biosg/GCATTCCTGCTGAACCGCTCTTCCGATCT | GAGTGCTT | TTTTTTTTTTTTTTTTTTTTVN |
T20.id10 | T20VN.BC8.PE_R.Indx10 | /5Biosg/GCATTCCTGCTGAACCGCTCTTCCGATCTATACGCCATTTTTTTTTTTTTTTTTTTTVN | /5Biosg/GCATTCCTGCTGAACCGCTCTTCCGATCT | ATACGCCA | TTTTTTTTTTTTTTTTTTTTVN |
T20.id11 | T20VN.BC8.PE_R.Indx11 | /5Biosg/GCATTCCTGCTGAACCGCTCTTCCGATCTATGCGTTATTTTTTTTTTTTTTTTTTTTVN | /5Biosg/GCATTCCTGCTGAACCGCTCTTCCGATCT | ATGCGTTA | TTTTTTTTTTTTTTTTTTTTVN |
T20.id12 | T20VN.BC8.PE_R.Indx12 | /5Biosg/GCATTCCTGCTGAACCGCTCTTCCGATCTTCACCCTTTTTTTTTTTTTTTTTTTTTTVN | /5Biosg/GCATTCCTGCTGAACCGCTCTTCCGATCT | TCACCCTT | TTTTTTTTTTTTTTTTTTTTVN |
T20.id13 | T20V.BC8.PE_R.Indx13 | /5Biosg/GCATTCCTGCTGAACCGCTCTTCCGATCTCCAACCGCTTTTTTTTTTTTTTTTTTTTV | /5Biosg/GCATTCCTGCTGAACCGCTCTTCCGATCT | CCAACCGC | TTTTTTTTTTTTTTTTTTTTV |
T20.id14 | T20V.BC8.PE_R.Indx14 | /5Biosg/GCATTCCTGCTGAACCGCTCTTCCGATCTGGATTACCTTTTTTTTTTTTTTTTTTTTV | /5Biosg/GCATTCCTGCTGAACCGCTCTTCCGATCT | GGATTACC | TTTTTTTTTTTTTTTTTTTTV |
T20.id15 | T20V.BC8.PE_R.Indx15 | /5Biosg/GCATTCCTGCTGAACCGCTCTTCCGATCTCAGAGACATTTTTTTTTTTTTTTTTTTTV | /5Biosg/GCATTCCTGCTGAACCGCTCTTCCGATCT | CAGAGACA | TTTTTTTTTTTTTTTTTTTTV |
T20.id16 | T20V.BC8.PE_R.Indx16 | /5Biosg/GCATTCCTGCTGAACCGCTCTTCCGATCTCAATAGTGTTTTTTTTTTTTTTTTTTTTV | /5Biosg/GCATTCCTGCTGAACCGCTCTTCCGATCT | CAATAGTG | TTTTTTTTTTTTTTTTTTTTV |
T20.id17 | T20V.BC8.PE_R.Indx17 | /5Biosg/GCATTCCTGCTGAACCGCTCTTCCGATCTGGTGGCCATTTTTTTTTTTTTTTTTTTTV | /5Biosg/GCATTCCTGCTGAACCGCTCTTCCGATCT | GGTGGCCA | TTTTTTTTTTTTTTTTTTTTV |
T20.id18 | T20V.BC8.PE_R.Indx18 | /5Biosg/GCATTCCTGCTGAACCGCTCTTCCGATCTGGAGGGAGTTTTTTTTTTTTTTTTTTTTV | /5Biosg/GCATTCCTGCTGAACCGCTCTTCCGATCT | GGAGGGAG | TTTTTTTTTTTTTTTTTTTTV |
T20.id19 | T20V.BC8.PE_R.Indx19 | /5Biosg/GCATTCCTGCTGAACCGCTCTTCCGATCTGTTGTCCTTTTTTTTTTTTTTTTTTTTTV | /5Biosg/GCATTCCTGCTGAACCGCTCTTCCGATCT | GTTGTCCT | TTTTTTTTTTTTTTTTTTTTV |
T20.id20 | T20V.BC8.PE_R.Indx20 | /5Biosg/GCATTCCTGCTGAACCGCTCTTCCGATCTTTCGACCTTTTTTTTTTTTTTTTTTTTTV | /5Biosg/GCATTCCTGCTGAACCGCTCTTCCGATCT | TTCGACCT | TTTTTTTTTTTTTTTTTTTTV |
T20.id21 | T20V.BC8.PE_R.Indx21 | /5Biosg/GCATTCCTGCTGAACCGCTCTTCCGATCTGCTTATTGTTTTTTTTTTTTTTTTTTTTV | /5Biosg/GCATTCCTGCTGAACCGCTCTTCCGATCT | GCTTATTG | TTTTTTTTTTTTTTTTTTTTV |
T20.id22 | T20V.BC8.PE_R.Indx22 | /5Biosg/GCATTCCTGCTGAACCGCTCTTCCGATCTCAACCTTTTTTTTTTTTTTTTTTTTTTTV | /5Biosg/GCATTCCTGCTGAACCGCTCTTCCGATCT | CAACCTTT | TTTTTTTTTTTTTTTTTTTTV |
T20.id23 | T20V.BC8.PE_R.Indx23 | /5Biosg/GCATTCCTGCTGAACCGCTCTTCCGATCTCTTAGGTATTTTTTTTTTTTTTTTTTTTV | /5Biosg/GCATTCCTGCTGAACCGCTCTTCCGATCT | CTTAGGTA | TTTTTTTTTTTTTTTTTTTTV |
T20.id24 | T20V.BC8.PE_R.Indx24 | /5Biosg/GCATTCCTGCTGAACCGCTCTTCCGATCTGCTCACCATTTTTTTTTTTTTTTTTTTTV | /5Biosg/GCATTCCTGCTGAACCGCTCTTCCGATCT | GCTCACCA | TTTTTTTTTTTTTTTTTTTTV |
N2.id73 | PCR_R.N2Ind73 | CAAGCAGAAGACGGCATACGAGATGAGTCCCTCGGCATTCCTGCTGAACCGCTCTT | CAAGCAGAAGACGGCATACGAGAT | GAGTCC | CTCGGCATTCCTGCTGAACCGCTCTT |
N2.id74 | PCR_R.N2Ind74 | CAAGCAGAAGACGGCATACGAGATTCGCCACTCGGCATTCCTGCTGAACCGCTCTT | CAAGCAGAAGACGGCATACGAGAT | TCGCCA | CTCGGCATTCCTGCTGAACCGCTCTT |
N2.id75 | PCR_R.N2Ind75 | CAAGCAGAAGACGGCATACGAGATAAGTCGCTCGGCATTCCTGCTGAACCGCTCTT | CAAGCAGAAGACGGCATACGAGAT | AAGTCG | CTCGGCATTCCTGCTGAACCGCTCTT |
N2.id76 | PCR_R.N2Ind76 | CAAGCAGAAGACGGCATACGAGATAATAGGCTCGGCATTCCTGCTGAACCGCTCTT | CAAGCAGAAGACGGCATACGAGAT | AATAGG | CTCGGCATTCCTGCTGAACCGCTCTT |
N2.id77 | PCR_R.N2Ind77 | CAAGCAGAAGACGGCATACGAGATACCCGTCTCGGCATTCCTGCTGAACCGCTCTT | CAAGCAGAAGACGGCATACGAGAT | ACCCGT | CTCGGCATTCCTGCTGAACCGCTCTT |
N2.id78 | PCR_R.N2Ind78 | CAAGCAGAAGACGGCATACGAGATAACACGCTCGGCATTCCTGCTGAACCGCTCTT | CAAGCAGAAGACGGCATACGAGAT | AACACG | CTCGGCATTCCTGCTGAACCGCTCTT |
N2.id79 | PCR_R.N2Ind79 | CAAGCAGAAGACGGCATACGAGATGCTTGGCTCGGCATTCCTGCTGAACCGCTCTT | CAAGCAGAAGACGGCATACGAGAT | GCTTGG | CTCGGCATTCCTGCTGAACCGCTCTT |
N2.id80 | PCR_R.N2Ind80 | CAAGCAGAAGACGGCATACGAGATTTACCACTCGGCATTCCTGCTGAACCGCTCTT | CAAGCAGAAGACGGCATACGAGAT | TTACCA | CTCGGCATTCCTGCTGAACCGCTCTT |
N2.id81 | PCR_R.N2Ind81 | CAAGCAGAAGACGGCATACGAGATCCAGGTCTCGGCATTCCTGCTGAACCGCTCTT | CAAGCAGAAGACGGCATACGAGAT | CCAGGT | CTCGGCATTCCTGCTGAACCGCTCTT |
N2.id82 | PCR_R.N2Ind82 | CAAGCAGAAGACGGCATACGAGATCGTTTGCTCGGCATTCCTGCTGAACCGCTCTT | CAAGCAGAAGACGGCATACGAGAT | CGTTTG | CTCGGCATTCCTGCTGAACCGCTCTT |
N2.id83 | PCR_R.N2Ind83 | CAAGCAGAAGACGGCATACGAGATGACCACCTCGGCATTCCTGCTGAACCGCTCTT | CAAGCAGAAGACGGCATACGAGAT | GACCAC | CTCGGCATTCCTGCTGAACCGCTCTT |
N2.id84 | PCR_R.N2Ind84 | CAAGCAGAAGACGGCATACGAGATACAAGACTCGGCATTCCTGCTGAACCGCTCTT | CAAGCAGAAGACGGCATACGAGAT | ACAAGA | CTCGGCATTCCTGCTGAACCGCTCTT |
N2.id85 | PCR_R.N2Ind85 | CAAGCAGAAGACGGCATACGAGATACCGCACTCGGCATTCCTGCTGAACCGCTCTT | CAAGCAGAAGACGGCATACGAGAT | ACCGCA | CTCGGCATTCCTGCTGAACCGCTCTT |
N2.id86 | PCR_R.N2Ind86 | CAAGCAGAAGACGGCATACGAGATTGGGTACTCGGCATTCCTGCTGAACCGCTCTT | CAAGCAGAAGACGGCATACGAGAT | TGGGTA | CTCGGCATTCCTGCTGAACCGCTCTT |
N2.id87 | PCR_R.N2Ind87 | CAAGCAGAAGACGGCATACGAGATATTCCGCTCGGCATTCCTGCTGAACCGCTCTT | CAAGCAGAAGACGGCATACGAGAT | ATTCCG | CTCGGCATTCCTGCTGAACCGCTCTT |
N2.id88 | PCR_R.N2Ind88 | CAAGCAGAAGACGGCATACGAGATGAATGTCTCGGCATTCCTGCTGAACCGCTCTT | CAAGCAGAAGACGGCATACGAGAT | GAATGT | CTCGGCATTCCTGCTGAACCGCTCTT |
N2.id89 | PCR_R.N2Ind89 | CAAGCAGAAGACGGCATACGAGATGCTGATCTCGGCATTCCTGCTGAACCGCTCTT | CAAGCAGAAGACGGCATACGAGAT | GCTGAT | CTCGGCATTCCTGCTGAACCGCTCTT |
N2.id90 | PCR_R.N2Ind90 | CAAGCAGAAGACGGCATACGAGATAGTGCTCTCGGCATTCCTGCTGAACCGCTCTT | CAAGCAGAAGACGGCATACGAGAT | AGTGCT | CTCGGCATTCCTGCTGAACCGCTCTT |
N2.id91 | PCR_R.N2Ind91 | CAAGCAGAAGACGGCATACGAGATCAGGGACTCGGCATTCCTGCTGAACCGCTCTT | CAAGCAGAAGACGGCATACGAGAT | CAGGGA | CTCGGCATTCCTGCTGAACCGCTCTT |
N2.id92 | PCR_R.N2Ind92 | CAAGCAGAAGACGGCATACGAGATCATGCGCTCGGCATTCCTGCTGAACCGCTCTT | CAAGCAGAAGACGGCATACGAGAT | CATGCG | CTCGGCATTCCTGCTGAACCGCTCTT |
N2.id93 | PCR_R.N2Ind93 | CAAGCAGAAGACGGCATACGAGATTGCCTACTCGGCATTCCTGCTGAACCGCTCTT | CAAGCAGAAGACGGCATACGAGAT | TGCCTA | CTCGGCATTCCTGCTGAACCGCTCTT |
N2.id94 | PCR_R.N2Ind94 | CAAGCAGAAGACGGCATACGAGATCTATACCTCGGCATTCCTGCTGAACCGCTCTT | CAAGCAGAAGACGGCATACGAGAT | CTATAC | CTCGGCATTCCTGCTGAACCGCTCTT |
N2.id95 | PCR_R.N2Ind95 | CAAGCAGAAGACGGCATACGAGATCCGAGTCTCGGCATTCCTGCTGAACCGCTCTT | CAAGCAGAAGACGGCATACGAGAT | CCGAGT | CTCGGCATTCCTGCTGAACCGCTCTT |
N2.id96 | PCR_R.N2Ind96 | CAAGCAGAAGACGGCATACGAGATACCTGCCTCGGCATTCCTGCTGAACCGCTCTT | CAAGCAGAAGACGGCATACGAGAT | ACCTGC | CTCGGCATTCCTGCTGAACCGCTCTT |
Sequencing primers[edit]
totoRNAseq Read 1: [1]
CACCGAGATCTACACTCTTTCCCTACACGACGTT 34nt Tm=64(IDT) Tm=71.72(Kun's) correspond to TSO.idXX primers used for 5'end
totoRNAseq Read 1 v2:
CCACCGAGATCTACACTCTTTCCCTACACGACG 33nt Tm=64.5(IDT) Tm=72.04(Kun's) correspond to TSO.r02/TSO.r04 primers used for 5'end