After ligation,I found that my colony on amplicilin plate have not insert geneligation digestion
Posted 17 January 2013 - 09:31 AM
My problems are....1. How did "insert gene" lost?(while there are colony on amplicilin primary plate and weak band in gel)
2. Why? molecular weight band of plasmid are not about 5420-5900bp but are 3200bp?
I don't understand T_T reply please.I'm worry.
Thank you very much
Posted 17 January 2013 - 11:45 AM
If you only have a few colonies from a transformation, and your positive control transformation has lots, then there is probably something wrong with your ligation reaction. You should also try to transform appropriate ligation controls (e.g. cut vector +ligase, cut vector alone).
Posted 13 December 2016 - 05:56 PM
I was wondering if you have pETDuet vector that you wouldn't mind sharing with me.
Looking forward to hearing from you
Posted 13 December 2016 - 07:32 PM
@Ribozome - the date on the original post was 2013, so it is unlikely that the original poster is still monitoring this post. Even if they were monitoring, pETDuet seems to be a commercial plasmid, so it is very unlikely that the OP could share it with you as these come with licences restricting use to the purchaser.