A novel vector for construction of cDNA library | Zendy

V.I. Fedchenko | Zendy; A.A. Kaloshin | Zendy; A. E. Medvedev | Zendy

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A novel vector for construction of cDNA library

Author(s) -

V.I. Fedchenko,

A.A. Kaloshin,

A. E. Medvedev

Publication year - 2010

Publication title -

biomeditsinskaya khimiya

Language(s) - English

Resource type - Journals

eISSN - 2310-6972

pISSN - 2310-6905

DOI - 10.18097/pbmc20105603329

Subject(s) - complementary dna , cdna library , primer (cosmetics) , polyadenylation , oligonucleotide , recombinant dna , microbiology and biotechnology , vector (molecular biology) , biology , rapid amplification of cdna ends , computational biology , dna , dna sequencing , genetics , messenger rna , chemistry , gene , molecular cloning , organic chemistry

A new original vector pEM-(dT)40(f+) has been prepared. It can be used for cDNA library construction from polyadenylated mRNA, isolated from various sources. The pGEM-(dT)40f(+) is initially transformed into single stranded and then into a linear form and its (dT)40 tail at 3'-end is used as the vector-primer for synthesis of the first strand cDNA. The use of a synthetic oligonucleotide complementary to the vector and recombinant DNA results in vector cyclization and synthesis of the second strand cDNA. This approach significantly simplifies cDNA library construction, it does not require PCR reaction (which can induce artifact mutations in cDNA sequences) and restrictase treatment.

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