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

From ZhangLabWiki
Jump to navigation Jump to search
>Jie deng
No edit summary
>Jie deng
No edit summary
 
(2 intermediate revisions by the same user not shown)
Line 1: Line 1:
==MMP9 ASE of PGP1F and PGP1L==
==MMP9 ASE of PGP1F and PGP1L==
The cDNA of PGP1F and PGP1L: refer to LabNotes [http://genome-tech.ucsd.edu/LabNotes/index.php/Jie:LabNotes/CpgSeq/2009-7-25]
===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         
Line 15: Line 17:
[[Image:20090727_PGP1F_PGP1L_MMP9_ASE.jpg|300px]]20090727_PGP1F_PGP1L_MMP9_ASE
[[Image:20090727_PGP1F_PGP1L_MMP9_ASE.jpg|300px]]20090727_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.
  The sanger sequencing results did not show ASE of MMP9 in two cell lines.

Latest revision as of 18:49, 28 July 2009

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.