I have screened 10 colonies out of 22. But neither get insert release nor the PCR amplification for the desired gene. What should I do, should I further screen the colonies?
Sounds like you got no amplification? If so, you should include any kind of positive control. Besides that, screening a 12 more isnt that much workload and is advisable. If I want to screen big numbers I do also pooled screenings where I include material from 5 to 8 colonies in one pcr to check if at least one in the badge is positive. If not I tested 8 in a single pcr. If one is positive its only 8 more pcrs to check which one. This way I tested once 170 colonies of which one was positive. So yes doing the big screening can be beneficial.
I agree but with so few colonies - ideally you want ~ 100 - i would re ligate and transform or at the very least re transform and screen some more
I have to disagree with the previous advice.
Screen the rest of your colonies. It's not much time to set up liquid cultures, do a quick plasmid prep, set up a digest, and run them out on a gel. If you set up the liquid cultures late today, you can do the rest tomorrow by lunch.
You only need one that worked the way you want. You'll know by the end of the day tomorrow if you need to repeat the transformation.
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