Hosuk:FISSEQ Primer information: Difference between revisions
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>Hosuki78 (Created page with "*FISSEQ_RT : RT primer for 1st Rolony fabrication **/5Phos/TCTCGGGAACGCTGAAGANNNNNN *FISSEQ_RCA : RCA primer for 1st Rolony fabrication **TCTTCAGCGTTCCCGA*G*A *FISSEQ_Adpt...") |
>Hosuki78 No edit summary |
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===Primers=== | |||
*FISSEQ_RT : RT primer for 1st Rolony fabrication | *FISSEQ_RT : RT primer for 1st Rolony fabrication | ||
**/5Phos/TCTCGGGAACGCTGAAGANNNNNN | **/5Phos/TCTCGGGAACGCTGAAGANNNNNN | ||
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*FISSEQ_ppRCA : RCA primer for 2nd Rolony fab. from padlock probes | |||
**GATATCGGGAAGCTGA*A*G | |||
===Padlock Probes==== | |||
*ppACTB | |||
**/5Phos/CTGTGCTCGCGGGGCGCTTCAGCTTCCCGATATCCGACGGACGTATCGGTAGTCGCAACGCAGGCAAAGGCGAGGCT | |||
*ppRAB7A | |||
**/5Phos/GAAGCGAGAAGGTCCAAGTTCTGCTTCAGCTTCCCGATATCCGACGGGTCTTGCGTGCGATACGGAGTAGAGGAGACTAAACGGAGGACA | |||
*ppMALAT1 | |||
**/5Phos/TTTCTGCCTTTACTTATCAATTCCTTCAGCTTCCCGATATCCGACGGTCTACTTCGTCGCGTCAGACCAAATGGAGGTATGACATATAATCT | |||
===Detection Probes=== | |||
*FISSEQ_Adpt : Cy3 labeled probe for 1st Rolony detection | *FISSEQ_Adpt : Cy3 labeled probe for 1st Rolony detection | ||
**/5Cy3/TCTCGGGAACGCTGAAGA | **/5Cy3/TCTCGGGAACGCTGAAGA | ||
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*FISSEQ_AdptCy5 : Cy5 labeled probe for 1st Rolony detection | *FISSEQ_AdptCy5 : Cy5 labeled probe for 1st Rolony detection | ||
**/5Cy5/TCTCGGGAACGCTGAAGA | **/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 ppMALAL1 | |||
**Cy3-TCTACTTCGTCGCGTCAGACCA | |||
Revision as of 18:50, 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=
- ppACTB
- /5Phos/CTGTGCTCGCGGGGCGCTTCAGCTTCCCGATATCCGACGGACGTATCGGTAGTCGCAACGCAGGCAAAGGCGAGGCT
- ppRAB7A
- /5Phos/GAAGCGAGAAGGTCCAAGTTCTGCTTCAGCTTCCCGATATCCGACGGGTCTTGCGTGCGATACGGAGTAGAGGAGACTAAACGGAGGACA
- ppMALAT1
- /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 ppMALAL1
- Cy3-TCTACTTCGTCGCGTCAGACCA
- RT-dT50 : Cy3 labeled probe for mRNA detection
- /5Cy3/TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTT
- RT-Adapt : Cy5 labeled probe for cDNA after RT
- /5Cy5/TCTTCAGCGTTCCCTAGA