Query on the Barnes paper/ Long range PCR
annag at vifp.monash.edu.au
Wed Apr 6 20:27:57 EST 1994
In the Barnes paper, he uses up to 10ng of template DNA equivalent to 7ng of the 35kb target sequence. This is a large amount of template DNA to start with especially compared with short range PCR. Eg, we use 1ng of human DNA to synthesize a 200 bp fragment ie, 1/10,000th of a pg of target DNA.
Is the method of Barnes all that efficient at amplifying DNA or is it only slightly amplifying the target DNA you start with?
The efficiency of amplifying long range PCR products doesn't seem to have been greatly improved by the use of the enzyme combinations.
Unless the efficiency of the system is dramaticaly improved it doesn't seem to be practical for use with genomic DNA.
Interested in your comments!
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