Molecular Cloning: A Laboratory Manual, Book 2 |
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Page 55
1985 ) or Sequenase " ( Schena 1989 ) , which cannot readily remove the
hybridized mutagenic primer from its template . The hybrid formed between the
template and the 3 ' terminus of the oligonucleotide is sufficiently stable to allow
priming ...
1985 ) or Sequenase " ( Schena 1989 ) , which cannot readily remove the
hybridized mutagenic primer from its template . The hybrid formed between the
template and the 3 ' terminus of the oligonucleotide is sufficiently stable to allow
priming ...
Page 57
7 ) , two oligonucleotides — the phosphorylated mutagenic oligonucleotide and a
universal sequencing primer ( which need not be phosphorylated ) — are mixed
in a 10 - to 50 - fold molar excess with the single - stranded template DNA in a ...
7 ) , two oligonucleotides — the phosphorylated mutagenic oligonucleotide and a
universal sequencing primer ( which need not be phosphorylated ) — are mixed
in a 10 - to 50 - fold molar excess with the single - stranded template DNA in a ...
Page 63
The mutagenic oligonucleotide should be designed as described on pages 15 .
54 – 15 . 56 and should be complementary to the strand of the target DNA that is
packaged in bacteriophage M13 particles ( the ( + ) strand ) . Before use in site ...
The mutagenic oligonucleotide should be designed as described on pages 15 .
54 – 15 . 56 and should be complementary to the strand of the target DNA that is
packaged in bacteriophage M13 particles ( the ( + ) strand ) . Before use in site ...
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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 |
Copyright | |
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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