pcr for gene fusion - (Jun/13/2007 )
hi
I want to fuse a 1kb DNA fragment with a 600 bp one and clone this chimeric gene in a vector. Both the fragments are pcr products. I have heard that instead of using the conventional restriction digestion ligation method the two fragments can be fused together by a pcr based strategy. Can anyone suggest how to go about this?
-research scholar-
QUOTE (research scholar @ Jun 13 2007, 02:18 AM)
hi
I want to fuse a 1kb DNA fragment with a 600 bp one and clone this chimeric gene in a vector. Both the fragments are pcr products. I have heard that instead of using the conventional restriction digestion ligation method the two fragments can be fused together by a pcr based strategy. Can anyone suggest how to go about this?
I want to fuse a 1kb DNA fragment with a 600 bp one and clone this chimeric gene in a vector. Both the fragments are pcr products. I have heard that instead of using the conventional restriction digestion ligation method the two fragments can be fused together by a pcr based strategy. Can anyone suggest how to go about this?
Yes, its very easy..
you can use the Overlap Extension PCR.. i fused 900 bp pcr product with 200 bp.. lol
last month (May07) published a good review in Nature Methods..
http://labclone.blogspot.com/2007/06/overl...ension-pcr.html
http://www.nature.com/nmeth/journal/v4/n5/...eth0507-455.pdf
if you don't understand any thing.. plzz feel free to ask.
Amit
University of Oxford
-akslal-
QUOTE (akslal @ Jun 13 2007, 03:56 AM)
QUOTE (research scholar @ Jun 13 2007, 02:18 AM)
hi
I want to fuse a 1kb DNA fragment with a 600 bp one and clone this chimeric gene in a vector. Both the fragments are pcr products. I have heard that instead of using the conventional restriction digestion ligation method the two fragments can be fused together by a pcr based strategy. Can anyone suggest how to go about this?
I want to fuse a 1kb DNA fragment with a 600 bp one and clone this chimeric gene in a vector. Both the fragments are pcr products. I have heard that instead of using the conventional restriction digestion ligation method the two fragments can be fused together by a pcr based strategy. Can anyone suggest how to go about this?
Yes, its very easy..
you can use the Overlap Extension PCR.. i fused 900 bp pcr product with 200 bp.. lol
last month (May07) published a good review in Nature Methods..
http://labclone.blogspot.com/2007/06/overl...ension-pcr.html
http://www.nature.com/nmeth/journal/v4/n5/...eth0507-455.pdf
if you don't understand any thing.. plzz feel free to ask.
Amit
University of Oxford
-research scholar-
thanks a lot Amit
-research scholar-