Open AccessConditions for Using DNA Polymerase I as an RNA-Dependent DNA Polymerase
Author(s) -
Subhash C. Gulati,
Daniel Kacian,
S. Spiegelman
Publication year - 1974
Publication title -
proceedings of the national academy of sciences
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 5.011
H-Index - 771
eISSN - 1091-6490
pISSN - 0027-8424
DOI - 10.1073/pnas.71.4.1035
Subject(s) - polymerase , dna clamp , dna polymerase , dna , dna polymerase ii , dna polymerase i , microbiology and biotechnology , biology , rna polymerase , rna , nucleic acid thermodynamics , chemistry , biochemistry , reverse transcriptase , gene , base sequence
Conditions are described for using Escherichia coli DNA polymerase I for synthesizing complementary DNA copies of natural RNA molecules, which are suitable for use in hybridization experiments. The molar ratio of enzyme to template is critical; below a certain level, synthesis is not observed. Hybrids formed with the complementary DNA are of comparable specificity and stability to those formed with complementary DNAs synthesized by viral RNA-directed DNA polymerase. Synthesis of dA-dT polymers, a common occurrence with this enzyme, can be eliminated by including distamycin in the reaction mixture.
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