lic-primer-design.gif
by Nick on February 18, 2008
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Nick Oswald
Nick is a molecular biologist who grazes in the field of biocatalysis by day and cooks up Bitesize Bio by night.
Nick is a molecular biologist who grazes in the field of biocatalysis by day and cooks up Bitesize Bio by night.
February 19th, 2008 at 10:56 am edit
Hm. How about this description:
Design your primers pF and pR with primer3 as usual, write them down in the usual 5′-3′ direction. Before ordering, you have to add some basepairs:
Take 15 bp from the left side (pleft) and 15 bp from the right side (pright) of your restriction site in the plasmid. Then:
a) LIC primer left: pleft pF
b) LIC primer right: reverse complement(pright) pR
The reverse overlap has to be reverse complemented. Then Simply add your overlapping regions 5′ to each primer.
Hm. Don’t know if this is easier to understand…