genome walking - (Jun/29/2007 )

Hi all,

I got clontech genome walking kit recently, i tried to amplify the control library with specified primers& conditions using platinum Pfx (invitrogen) with no success. i tried different conditions but failed.

Can any one suggest me whether i can use DNA polymerases other than clontech Advantage 2 PCR kit(recommended by clontech), if so where i am going wrong.

thanks in advance for your suggestions.

regards

gene_tag

If it's a new kit and you tested with the controls and got nothing.... then you can really ask for a new kit because it might be a bad batch.

Normally, kit manual will not recommend using 3rd party material. I'm not sure about your case but if you plan to use a 3rd party enzyme, there should be modifications such as buffer usage, extension programs etc. and also optimizations. As for components that came with the kit, the manufacturer normally had optimized the conditions (and with a troubleshooting guide).