Molecular Cloning: A Laboratory Manual, Book 2 |
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Page 80
SP6 promoter Polycloning Site Hindili Sall BamHI Smal BstEll EcoRI In PSP64CS
, the Hindill site in the polycloning site is downstream from the SP6 promoter . In
PSP65CS , the EcoRI site in the polycloning site is downstream from the SP6 ...
SP6 promoter Polycloning Site Hindili Sall BamHI Smal BstEll EcoRI In PSP64CS
, the Hindill site in the polycloning site is downstream from the SP6 promoter . In
PSP65CS , the EcoRI site in the polycloning site is downstream from the SP6 ...
Page 81
... and PSP65CS ) contain a polycloning site with. 5 ' PSP64CS Tth 1111 Tth
1111 Smal . ... 11B To use PSP64CS / PSP65CS , the target DNA is cloned into
the central Smal site . After cleavage of the resulting recombinant plasmid with ...
... and PSP65CS ) contain a polycloning site with. 5 ' PSP64CS Tth 1111 Tth
1111 Smal . ... 11B To use PSP64CS / PSP65CS , the target DNA is cloned into
the central Smal site . After cleavage of the resulting recombinant plasmid with ...
Page 82
11 , a recombinant plasmid carrying the target at the central Smal site is cleaved
with Tth1111 , generating two fragments of DNA . Each of the four termini carries
a different protruding base . By choosing the appropriate [ P ] dNTP , it is ...
11 , a recombinant plasmid carrying the target at the central Smal site is cleaved
with Tth1111 , generating two fragments of DNA . Each of the four termini carries
a different protruding base . By choosing the appropriate [ P ] dNTP , it 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