Molecular Cloning: A Laboratory Manual, Book 2 |
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Page 31
These partners ( 5 ' AGCGCTC ) bind to complementary sequences of the primer
- adapter , forming small double - stranded segments that can establish
phosphodiester bonds with both strands of the bacteriophage vector . After
attachment of ...
These partners ( 5 ' AGCGCTC ) bind to complementary sequences of the primer
- adapter , forming small double - stranded segments that can establish
phosphodiester bonds with both strands of the bacteriophage vector . After
attachment of ...
Page 20
Primer : Template Ratios and Nucleotide Concentrations The size , specific
activity , and yield of the radioactive strand synthesized by the Klenow fragment
of E . coli DNA polymerase I are affected by the relative concentrations of primer ...
Primer : Template Ratios and Nucleotide Concentrations The size , specific
activity , and yield of the radioactive strand synthesized by the Klenow fragment
of E . coli DNA polymerase I are affected by the relative concentrations of primer ...
Page 76
... dNTPs , primer , radioactive precursor ) omitted from reaction Saran Wrap not
removed from gel after drying No template DNA Primer - binding site deleted from
template High concentrations of EDTA present Incorrect primer used Check that ...
... dNTPs , primer , radioactive precursor ) omitted from reaction Saran Wrap not
removed from gel after drying No template DNA Primer - binding site deleted from
template High concentrations of EDTA present Incorrect primer used Check that ...
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Contents
TEST LIGATION 3 29 | 3 |
SYNTHESIS OF THE FIRST STRAND OF cDNA 8 | 11 |
FRAGMENT OF E coli DNA POLYMERASE I 13 59 | 14 |
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Common terms and phrases
acid activity agarose aliquots allow amount amplification appropriate approximately bacteriophage bacteriophage T4 base buffer carried cDNA cells centrifugation Chapter chromatography cleaved clones colonies concentration constructed containing deletions denatured described determine digestion DNA polymerase dNTPs double-stranded efficiency electrophoresis et al ethanol expression Figure filters fragment gene genomic hybridization increase Incubate inserted interest isolated labeled length libraries ligation linear linkers method microfuge minutes mixture molecules mRNA mutagenesis mutagenic mutations Nucleic nucleotides obtained oligonucleotide original plaques plasmid plasmid DNA plate pool position possible prepared presence primer probe problem protein purified radiolabeled reaction recombinant region remove restriction enzyme room temperature samples screening sequence single single-stranded solution specific step Store strand synthesis target DNA target sequence template termini tion transfer transformants tube units usually vector volume