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Insert in frame - (Mar/02/2019 )

How to find out if the insert is inframe? I did expasy translate. Start and stop codon looks in place in frame 1 but not in frame 2 & 3? Is it ok to go ahead with the cloning? Please can someone help out.


In-frame compared to what? 


How did you clone the insert, and what are you trying to do?


If you used PCR to amplify your complete insert and then cloned this in using RE's outside the coding sequence, then you are most likely fine to go ahead. Frames 1, 2 and 3 are for comparing when your sequencing read doesn't necessarily start at the position you intended, so that you can see if there is an ORF there or not in any of the 3 possible reading frames.