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 > SMC1-mou-RV: GAGCTGTCCTCTCCTTG > > SMC1-mou-FWD: CTGTCATGGGTTTCCTG > > The amplified FS transcript is 162bp, Wt is 464bp Positive control cDNA 4T1 in mouse cDNA 9/25 at Tumor cDNA Expression Library 1-20-11 KW -20 C
 * the sequence:
 * >> 54.9 Salt adjusted Tm
 * [[file:SMC1fs PCR Settings 3-30-12.ppt]]

Notes from Andrey about PCR
 * 200 nM primer
 * 5 uL and 20 uL template
 * Use Taq or dynazyme

Results