Jie:LabNotes/CpgSeq/2009-7-27

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MMP9 ASE of PGP1F and PGP1L[edit]

The cDNA of PGP1F and PGP1L: refer to LabNotes [1]

PCR[edit]

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.