2 replies to this topic

[Ambinlab]

Dear All,

I have cloned an shRNA into lentiviral vector. I got some colonies (around 20). however I do not want to submit all of them for sequencing (too expensive!) and also do not know how to screen them to make sure which one has the insert.

Do you have any idea how I can find the right colony?

Thanks in advance

[pcrman]

The quick and dirty way is to do colony PCR, and then miniprep of those positive ones, and further verify by restriction digestion.

[Ambinlab]

Thanks
But the problem is that the template is so small that I cannot see it on a gel when I digest it...

any ideas?