Analysis of RNA by Primer Extension | Zendy

Michael R. Green | Zendy; Joseph Sambrook | Zendy

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Analysis of RNA by Primer Extension

Author(s) -

Michael R. Green,

Joseph Sambrook

Publication year - 2021

Publication title -

cold spring harbor protocols

Language(s) - English

Resource type - Journals

SCImago Journal Rank - 0.674

H-Index - 51

eISSN - 1940-3402

pISSN - 1559-6095

DOI - 10.1101/pdb.prot101840

Subject(s) - primer extension , primer (cosmetics) , microbiology and biotechnology , oligonucleotide , rna , messenger rna , reverse transcriptase , polyacrylamide gel electrophoresis , polynucleotide , biology , population , murine leukemia virus , chemistry , enzyme , dna , virus , biochemistry , genetics , gene , demography , sociology , organic chemistry

For mapping the 5′ termini of mRNA molecules, primer extension is the method of choice. A purified oligonucleotide is end-labeled using polynucleotide kinase. The probe and a population of mRNA are allowed to hybridize, and the primers and template are used to carry out reverse transcription using an enzyme cloned from the Moloney murine leukemia virus. The primer extension products are separated on a denaturing polyacrylamide gel and analyzed by radiography.

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