Hosuk:FISSEQ Primer information: Difference between revisions
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>Mzcai |
>Hosuki78 |
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===Padlock Probes=== | ===Padlock Probes=== | ||
====Structure==== | |||
*Probe: H1 + Common seq. for 2nd RCA + Common Linker + decoding probe seq. + H2 | |||
*Common seq. for 2nd RCA : CTTCAGCTTCCCGATATC | |||
*Common Linker : CGACGG | |||
====Padlock probes==== | |||
*ppACTB | *ppACTB | ||
**H1: | |||
**H2: | |||
**/5Phos/CTGTGCTCGCGGGGCGCTTCAGCTTCCCGATATCCGACGGACGTATCGGTAGTCGCAACGCAGGCAAAGGCGAGGCT | **/5Phos/CTGTGCTCGCGGGGCGCTTCAGCTTCCCGATATCCGACGGACGTATCGGTAGTCGCAACGCAGGCAAAGGCGAGGCT | ||
*ppRAB7A | *ppRAB7A | ||
**H1: | |||
**H2: | |||
**/5Phos/GAAGCGAGAAGGTCCAAGTTCTGCTTCAGCTTCCCGATATCCGACGGGTCTTGCGTGCGATACGGAGTAGAGGAGACTAAACGGAGGACA | **/5Phos/GAAGCGAGAAGGTCCAAGTTCTGCTTCAGCTTCCCGATATCCGACGGGTCTTGCGTGCGATACGGAGTAGAGGAGACTAAACGGAGGACA | ||
*ppMALAT1 | *ppMALAT1 | ||
**/5Phos/TTTCTGCCTTTACTTATCAATTCCTTCAGCTTCCCGATATCCGACGGTCTACTTCGTCGCGTCAGACCAAATGGAGGTATGACATATAATCT | **H1: TTTCTGCCTTTACTTATCAATTC | ||
**H2: AATGGAGGTATGACATATAATCT | |||
**/5Phos/TTTCTGCCTTTACTTATCAATTCCTTCAGCTTCCCGATATCCGACGGTCTACTTCGTCGCGTCAGACCAAATGGAGGTATGACATATAATCT | |||
===Detection Probes=== | ===Detection Probes=== |
Revision as of 19:00, 12 November 2013
Primers
- FISSEQ_RT : RT primer for 1st Rolony fabrication
- /5Phos/TCTCGGGAACGCTGAAGANNNNNN
- FISSEQ_RCA : RCA primer for 1st Rolony fabrication
- TCTTCAGCGTTCCCGA*G*A
- FISSEQ_ppRCA : RCA primer for 2nd Rolony fab. from padlock probes
- GATATCGGGAAGCTGA*A*G
Padlock Probes
Structure
- Probe: H1 + Common seq. for 2nd RCA + Common Linker + decoding probe seq. + H2
- Common seq. for 2nd RCA : CTTCAGCTTCCCGATATC
- Common Linker : CGACGG
Padlock probes
- ppACTB
- H1:
- H2:
- /5Phos/CTGTGCTCGCGGGGCGCTTCAGCTTCCCGATATCCGACGGACGTATCGGTAGTCGCAACGCAGGCAAAGGCGAGGCT
- ppRAB7A
- H1:
- H2:
- /5Phos/GAAGCGAGAAGGTCCAAGTTCTGCTTCAGCTTCCCGATATCCGACGGGTCTTGCGTGCGATACGGAGTAGAGGAGACTAAACGGAGGACA
- ppMALAT1
- H1: TTTCTGCCTTTACTTATCAATTC
- H2: AATGGAGGTATGACATATAATCT
- /5Phos/TTTCTGCCTTTACTTATCAATTCCTTCAGCTTCCCGATATCCGACGGTCTACTTCGTCGCGTCAGACCAAATGGAGGTATGACATATAATCT
Detection Probes
- FISSEQ_Adpt : Cy3 labeled probe for 1st Rolony detection
- /5Cy3/TCTCGGGAACGCTGAAGA
- FISSEQ_AdptCy5 : Cy5 labeled probe for 1st Rolony detection
- /5Cy5/TCTCGGGAACGCTGAAGA
- ACTB_1stRolonyFISH : ATTO488 labeled probe for ACTB on 1st Rolony
- GGCAAAGGCGAGGCT/3ATTO488N/
- RAB7A_1stRolonyFISH : ATTO550 labeled probe for RAB7A on 1st Rolony
- GAGGAGACTAAACGGAGGACA/3ATTO550N/
- Shifted FISSEQ_2ndRCAprimerDye : 2nd Rolony from all padlock probes
- GGATATCGGGAAGCTGAA/3ATTO488N/
- dcProbe0-Cy3 : for 2nd Rolony of ppACTB
- Cy3-ACGTATCGGTAGTCGCAACGCA
- dcProbe1-Cy3 : for 2nd Rolony of ppRAB7A
- Cy3-GTCTTGCGTGCGATACGGAGTA
- dcProbe2-Cy3 : for 2nd Rolony of ppMALAT1
- Cy3-TCTACTTCGTCGCGTCAGACCA
- RT-dT50 : Cy3 labeled probe for mRNA detection
- /5Cy3/TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTT
- RT-Adapt : Cy5 labeled probe for cDNA after RT
- /5Cy5/TCTTCAGCGTTCCCTAGA