Molecular Cloning: A Laboratory Manual, Book 2 |
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Page 16
However , if it is necessary to clone the 5 ' - terminal sequences of eukaryotic
mRNA with high efficiency , several more - difficult methods are available . For
example , after completion of first - strand synthesis , terminal transferase can be
used ...
However , if it is necessary to clone the 5 ' - terminal sequences of eukaryotic
mRNA with high efficiency , several more - difficult methods are available . For
example , after completion of first - strand synthesis , terminal transferase can be
used ...
Page 23
For optimal efficiency of cloning , the number of homopolymeric residues on the
plasmid and the cDNA should be approximately equal , with approximately 100
dA / dT residues or 20 dC / dG residues being added to each end of the DNAs ...
For optimal efficiency of cloning , the number of homopolymeric residues on the
plasmid and the cDNA should be approximately equal , with approximately 100
dA / dT residues or 20 dC / dG residues being added to each end of the DNAs ...
Page 78
The efficiency of transfection in ung * bacteria of bacteriophage M13 DNA that
contains uracil in one strand is greatly increased if the thyminecontaining strand
is covalently closed . This can only happen if the priming oligonucleotide ( s ) is ...
The efficiency of transfection in ung * bacteria of bacteriophage M13 DNA that
contains uracil in one strand is greatly increased if the thyminecontaining strand
is covalently closed . This can only happen if the priming oligonucleotide ( s ) is ...
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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