Dinh/NOTES/2010-7-3: Difference between revisions

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  -----------------------
  -----------------------
                     50ul
                     50ul
  Templates used: BJ-ips, HGPS-ips, HGPS72
  Templates used: HGPS-ips


  Program:  
  Program:  

Revision as of 16:39, 3 July 2010

Sanger sequencing of PCR product with probe linker

Primers

LinkerR-LMNA-F: GTT GGA GGC TCA TCG TTC CTA TTT TGG GCC TGA GTG GTC AGT C
LinkerL-LMNA-R: GAC CCG CCT GCA GGA TTT GGC AGG CAG ATG TTA TCG AGG TCC GAC
Product length: 373+23bp+25bp= 421bp
Map to position: chr1:156,108,195-156,108,590

PCR

Template (1-2ng/ul): 3ul
F primer (10uM)    : 1ul
R primer (10uM)    : 1ul
H20                : 20ul
Taq 2x MM          : 25ul
-----------------------
                   50ul
Templates used: HGPS-ips
Program: 
94C   2min
94C   30s
62C   40s
72C   40s
Go to 2 for 7x
94C   30s
72C   40s
Go to 7 for 23x
72C   3min
15C   forever
Purify by 0.9x volumes of Agencourt beads.
  • Quantification by Nanodrop


  • Premix for Genewiz sequencing:

Results