Haynes Lab:Notebook/David Barclay Undergrad Training/Plasmid Transformation/2013/10/27

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(Primers)
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{| width=600px
{| width=600px
|-
|-
-
|   || '''Roche Name''' || '''Left Primer''' || '''Right Primer'''
+
|   || '''Roche Name''' || '''Left Primer''' || '''Right Primer''' || '''UPL probe'''  
|-
|-
-
| PDX1 || NM_000209.3 || aagctcacgcgtggaaag || gccgtgagatgtacttgttgaa
+
| PDX1 || NM_000209.3 || gaaatccaccaaagctcacg || cgggttccgctgtgtaag || #51, cat.no. 04688481001
|-
|-
-
| MAFA || treated cells || MAFA
+
| MAFA || AB086960.1 || agcgagaagtgccaactcc || ttgtacaggtcccgctcttt || #39, cat.no. 04687973001
|-
|-
| GLP1R || NM_002062.3 || cacttggagctaattaaggtttgc || ccgtccagtctttctgctgt
| GLP1R || NM_002062.3 || cacttggagctaattaaggtttgc || ccgtccagtctttctgctgt

Revision as of 04:50, 7 November 2013

Haynes BioBrick Cloning Main project page
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UPLassay

  • This is a test run for RT-PCR training. Using gene targets identified in prior research to pancreas differentiation gene sequences.

Reaction List

REACTION LIST
  Template cDNA Gene Target
Rxn 1: treated cells PDX1
Rxn 2: treated cells MAFA
Rxn 3: treated cells GLP1R
Rxn 4: treated cells PCSK1
Rxn 5: treated cells IAPP
Rxn 6: treated cells KCNQ2
Rxn 7: treated cells GAPD (reference gene)
Rxn 8: untreated cells PDX1
Rxn 9: untreated cells MAFA
Rxn 10: untreated cells GLP1R
Rxn 11: untreated cells PCSK1
Rxn 12: untreated cells IAPP
Rxn 13: untreated cells KCNQ2
Rxn 14: untreated cells GAPD (reference gene)
Rxn 15: no template PDX1
Rxn 16: no template MAFA
Rxn 17: no template GLP1R
Rxn 18: no template PCSK1
Rxn 19: no template IAPP
Rxn 20: no template KCN2Q
Rxn 21: no template GAPD (reference gene)
  • Run 3 Each
  • 45 Reactions Run
  • Using Variation 2

Variation 2
Figure 1

  • Using as Reference, discount gene names on left side

Primers

PRIMERS LISt
  Roche Name Left Primer Right Primer UPL probe
PDX1 NM_000209.3 gaaatccaccaaagctcacg cgggttccgctgtgtaag #51, cat.no. 04688481001
MAFA AB086960.1 agcgagaagtgccaactcc ttgtacaggtcccgctcttt #39, cat.no. 04687973001
GLP1R NM_002062.3 cacttggagctaattaaggtttgc ccgtccagtctttctgctgt
PCSK1 treated cells PCSK1
IAPP treated cells IAPP
KCNQ2 treated cells KCNQ2
GAPD treated cells GAPD (reference gene)


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