Jie:LabNotes/CpgSeq/2009-7-27: Difference between revisions

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==MMP9 ASE of PGP1F and PGP1L==
==MMP9 ASE of PGP1F and PGP1L==
===PCR===
===PCR===
 
I designed the primer according to the alleotyping of PGP1. The target cover the rs13969(C/A) on exon.
  The primers: [[Media: 20090727_MMP9_ASE|MMP9_ASE_primer]]
  The primers: [[Media: 20090727_MMP9_ASE.xls|MMP9_ASE_primer]]
                                             x 4
                                             x 4
  2x iQ supermix                40          160         
  2x iQ supermix                40          160         

Revision as of 18:42, 28 July 2009

MMP9 ASE of PGP1F and PGP1L

PCR

I designed the primer according to the alleotyping of PGP1. The target cover the rs13969(C/A) on exon.
The primers: MMP9_ASE_primer
                                           x 4
2x iQ supermix                40           160         
Primer F (100uM)             0.4           1.6 each
Primer R (100uM)             0.4           1.6 each
cDNA samples as above:       0.1	  	
dH2O                          40           160
total                         80ul

96C 3min-> 45x (95C 30sec, 58C or 62C 1min, 72C 1min)-> 72C 5min.

File:20090727 PGP1F PGP1L MMP9 ASE.jpg20090727_PGP1F_PGP1L_MMP9_ASE

I sent PGP1F_62C and PGP1L_58C to do the sanger sequencing.
The sanger sequencing results did not show ASE of MMP9 in two cell lines.