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
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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