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Primers for Introduction of new restriction sites to a vector - (Mar/18/2012 )


I have designed primers to introduce new restriction sites to a vector to be able to introduce my cDNA, and I would appreciate it if anyone could check them and suggest any changes or confirm if it is correct.



LANDING SITE < MLUI > Kozak Fragment *GENE*

5’- GCGC GCCACC gcgggcagtagccgct -3’



3’- CTAG ccacatttattctacact -5’


My only comment is that you need to put the add-ons on the 5' end of the reverse primer always! You could copy and paste what you have written above into a primer ordering site and it would give you the correct oligo at the end, it's just that you have written the 5' and 3' at the wrong ends.


Thanks a lot for the reply...
What do you mean by the "add-ons"?


The restriction sites and other flanking bits.