SMC1fs PCR in 4T1 cDNA lysate pools 3-30-12

2015-01-13

azim58 - SMC1fs PCR in 4T1 cDNA lysate pools 3-30-12

experiment performed on 4-2-12

Many tumor cDNA lysate pools/spots other than the SMC1fs positive control
spot exhibited binding when anti-SMC1fs sera was applied to the array. We
are curious if some of these pools might contain clones with the SMC1fs
gene in them. We will do a PCR to test this. Note that there is a
possibility that the protein is not SMC1fs, but does have some type of
homologous sequence to it.

Primers from Shen: SMC1 Mus Jan 18 FW and RV at SMC1 9-20-10 KW 4 C

the sequence:

SMC1-mou-RV: GAGCTGTCCTCTCCTTG

>> 54.9 Salt adjusted Tm

SMC1-mou-FWD: CTGTCATGGGTTTCCTG

The amplified FS transcript is 162bp, Wt is 464bp

SMC1fs PCR Settings 3-30-12.ppt

Positive control cDNA 4T1 in mouse cDNA 9/25 at Tumor cDNA Expression
Library 1-20-11 KW -20 C

Notes from Andrey about PCR

200 nM primer
5 uL and 20 uL template
Use Taq or dynazyme

Results

SMC1fs PCR 3-30-12.svg
SMC1fs PCR 3-30-12.pdf