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

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  72C  3min
  72C  3min
  15C  forever
  15C  forever
==Results==
PAGE
[[File:Attaching linker to LMNA amplicons.jpg]]


  Purify by 0.9x volumes of Agencourt beads.
  Purify by 0.9x volumes of Agencourt beads.


* Quantification by Nanodrop
* Quantification by Nanodrop
* Premix for Genewiz sequencing:
==Results==

Revision as of 18:19, 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

Results

PAGE

File:Attaching linker to LMNA amplicons.jpg

Purify by 0.9x volumes of Agencourt beads.
  • Quantification by Nanodrop