Open AccessThe synthesis of high yields of full-length reverse transcripts of globin mRNA
Author(s) -
Eitan Friedman,
Michael Rosbash
Publication year - 1977
Publication title -
nucleic acids research
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 9.008
H-Index - 537
eISSN - 1362-4954
pISSN - 0305-1048
DOI - 10.1093/nar/4.10.3455
Subject(s) - complementary dna , reverse transcriptase , biology , microbiology and biotechnology , messenger rna , rapid amplification of cdna ends , cdna library , rna , biochemistry , gene , molecular cloning
Conditions have been determined under which reverse transcriptase catalyzes the synthesis of the high yields of full length complementary deoxyribonucleic acid (cDNA). These conditions depend not only on the cencentration of deoxynucleoside triphosphates (1) but also on the concentration of reverse transcriptase. An analysis of the kinetics of cDNA synthesis and the size of cDNA synthesized as a function of time under different conditions indicates that the mechanism of action of reverse transcriptase is partially distributive. This accounts for the necessity of a high enzyme concentration to obtain high yields of full length cDNA. Additional experiments indicate that the yield of cDNA is limited by the fact that the template mRNA is rapidly inactivated. This is most likely due to the fact that the product cDNA is hydrogen bonded to the template mRNA during synthesis.