Goodmorning, I was working with a cloning for some time now trying to get the eGFP out of a vector and add mcherry instead. I am at the final steps now and unfortunately found out that the forward primer used for the mcherry PCR was wrong. The mCherry translation will now start at th 2nd Met which is 9 aminoacids in the sequence (MVSKGEEDN is not translated). Has anybody had any experience with that??? Is there any chance the molecule's fluorecence will be OK??? I note that the mCherry will not be fused with any other protein.
Problem with mcherry cloning
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