Hi,
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.
Thanks.
Forward:
LANDING SITE [ MLUI ] Kozak Fragment [START] *GENE*
5’- GCGC [ACGCGT ] GCCACC [ATG] gcgggcagtagccgct -3’
Reverse:
LANDING SITE [BAMHI] [STOP] *GENE*
3’- CTAG [GGATCC] [CTA] ccacatttattctacact -5’
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Primers for Introduction of new restriction sites to a vector
Started by fabiola, Mar 17 2012 09:56 PM
vector restriction enzyme primers
3 replies to this topic
#1
Posted 17 March 2012 - 09:56 PM
#2
Posted 18 March 2012 - 04:41 PM
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.
- fabiola likes this
#3
Posted 18 March 2012 - 06:03 PM
Thanks a lot for the reply...
What do you mean by the "add-ons"?
What do you mean by the "add-ons"?
#4
Posted 19 March 2012 - 01:34 PM
The restriction sites and other flanking bits.
Also tagged with one or more of these keywords: vector, restriction enzyme, primers
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