Applying Random Primer and Overcoming Single Primer Amplification in PCR-based Chromosome Walking

Chromosome walking adopted a variety of PCR methods,including inverse PCR,ligation mediated PCR,arbitrarily primed PCR.Non-specific amplification by the universal single primer was common in these methods.The problems of single primer amplification and the preventive strategies were reviewed in this paper,primarily to design primers with only one binding site on the two end adaptors of the DNAs.Bubble-or Y-shaped,as well as single oligonucleotide adaptors with 3′ modifications of an amino group were commonly used.Mismatched primer and single primer PCR methods based on RAPD were also discussed.