Tips

Unknown gene detection procedures

Genomic DNA or cDNA preparation
DNA to be used as a template should be prepared by the usual method. Usually DNA can be isolated using the common method or kit. It is preferable to use the kit, rather than the manual to obtain quality DNA. Also, total RNA or plasmid (clone) may be used.

DNA Walking SpeedUp™ PCR
Three different PCR reactions will be conducted using (cDNA/genomic DNA) which you prepared. The first PCR uses DNA as a template, in which you have to design Target Specific Primer (TSP1) based on the known sequence. The design of TSP1 suitable to PCR conditions is very critical to generate the best results. The second and third PCRs use the previous PCR products and the third PCR product can be confirmed on agarose gel.