PCR Amplification of cDNA Libraries for Cloning and Screening

PCR amplification of cDNA libraries for cloning and screening.

Screening a cDNA library is a common practice for cloning genes of interest in many laboratories. Success of screening is largely dependent on the abundance of cDNAs representing genes of interest and the availability of suitable probes. The procedure involves several rounds of screening to obtain true-positive clones, which might take a week or longer. Typically, each round of screening requir...

Simplified protocol of solid-phase cDNA libraries for multiple PCR amplification.

Detection and evaluation of non-recombinants in cDNA libraries by multiple cloning region PCR.

Bacteriophages that are routinely used in cDNA libraries do not require any biological selection for forming plaques. Thus parental non-recombinant phages are always found in variable proportions together with recombinant ones in all cDNA libraries. The presence of non-recombinants in significant proportions dilutes the abundance of rare cDNA species and makes library screening difficult. If th...

Semi-multiplex PCR technique for screening of abundant transcripts during systematic sequencing of cDNA libraries.

The systematic sequencing of cDNA libraries is an efficient approach for the identification of new genes, but the presence of abundant mRNAs is often a major problem. This paper describes a very simple method of "semi-multiplex PCR" that allows specific identification of such abundant transcripts before DNA sequencing without using nonrepresentative subtracted libraries. The PCR utilizes a seri...

PCR-based Method for Screening Libraries

One of the fundamental techniques of molecular biology is the isolation of a rare clone from a complex mixture of DNA sequences contained within a library. The isolation of cDNA and genomic clones from highly complex libraries is often an early step in studies aimed at understanding basic biological processes as well as in applied biological research. In a typical genomic library with an averag...