deletion pcr - deletion pcr gene (Mar/13/2002 )
i research informations about deletion of gene by pcr. I have the method, but before i desire to know if anybody have problems with this process.
I am trying this technique right now, so I have no results no problems. I would like to know about your method so I could compare with mine.
there are some clever overlap pcr's u guys can do to get a gene deleted from a construct...
the complexity depends on hoe clever u r. idea is to do 2 sets of pcr's and then use the pcr prdt to do the 3rd pcr. and 3 pcr is the opverlap pcr where it will yield deleted insert or etc..
there are system u can buy like pfu/taq mixture that ui can use in TA cloning for the internmediate steps.
i found this technic on the web, in http://www.fhcrc.org/labs/gottschling/pcrko.sht i
idesire use the same primer, until you propose a best solution.