plan for pcr frameshift screen 05-30-2014d1050

2014-08-29

plan for pcr frameshift screen 05-30-2014d1050

email from Shen
https://mail.google.com/mail/u/0/?ui=2&shva=1#search/more+cDNA+samples/146441a4efae2e8c

Hi Kurt,
I just send one email. Something wrong with the gmail.
How about your job at Spain?
The cDNA set I mentioned is about 10 tumor cDNAs. You can ask Kelly for the details. Felicia should know where is the samples. Make more cDNA if some sample running out.
The strategy for the screen is:
First, screen the antibody positive cDNA. Each primer set I signed three positive cDNA samples.
Second, sequence all clear PCR products.
For the correct product, such as GF38, we should screen the set of cDNA I mentioned. However, you don't need to sequence these PCR. You just need to run all the PCR product with the sequenced PCR side by side.
For the sample has bright, clear and wrong size PCR product, you need to check the sequence to see if it is a different genefusion and encodes a FS peptide.
For the samples don't have PCR product. first, you need confirm the cDNA quality. then you can try different PCR strategy. If you still can not get PCR product, give up.

Let me know if you need anything more. Thanks!

Best,
Luhui Shen