PCR-based cDNA library construction: general cDNA libraries at the level of a few cells

PCR-based cDNA library construction: general cDNA libraries at the level of a few cells.

A procedure for the construction of general cDNA libraries is described which is based on the amplification of total cDNA in vitro. The first cDNA strand is synthesized from total RNA using an oligo(dT)-containing primer. After oligo(dG) tailing the total cDNA is amplified by PCR using two primers complementary to oligo(dA) and oligo(dG) ends of the cDNA. For insertion of the cDNA into a vector...

cDNA Microarray Strategy for the Construction of the cDNA Array Preparation of Subtraction Libraries Prescreening of Library Clones cDNA Microarray Production cDNA Microarray Composition

Subtractive hybridization and construction of cDNA libraries.

Normalization of a cDNA Library Cloned in λZAP by a Long PCR and cDNA Reassociation Procedure

250 BioTechniques Vol. 34, No. 2 (2003) be expected from an amplificationbased strategy. To our knowledge, this is the first time that such analysis has been conducted to assess the representation of a genomic library. In summary, we have developed a quick, facile protocol for constructing libraries from relatively small amounts of DNA. Using our method, we have shown that a representative, ran...

Construction of a normalized cDNA library by introduction of a semi-solid mRNA-cDNA hybridization system.

We report a novel procedure to construct a normalized (equalized) cDNA library. By introduction of the highly efficient self-hybridization system between a whole mRNA population and their corresponding cDNA immobilized on latex beads, which involves relatively simple manipulations, we were able to generate an mRNA population in which the copy number of abundant species was reduced while that of...