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.