Molecular Cloning: A Laboratory Manual, Book 2 |
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Page 14
SEQUENCING STRATEGIES Before beginning to sequence , it is important to
develop an overall strategy that takes into account the size of the region to be
sequenced , the accuracy of the sequence required , and the facilities that are ...
SEQUENCING STRATEGIES Before beginning to sequence , it is important to
develop an overall strategy that takes into account the size of the region to be
sequenced , the accuracy of the sequence required , and the facilities that are ...
Page 19
accumulated simultaneously from ordered regions of the target DNA . ... to design
the primer to be complementary to sequences that lie some distance behind the
advancing front , within a region of the gel that can be read with confidence . 2 .
accumulated simultaneously from ordered regions of the target DNA . ... to design
the primer to be complementary to sequences that lie some distance behind the
advancing front , within a region of the gel that can be read with confidence . 2 .
Page 32
Linker - scanning Mutagenesis The identification and localization of regulatory
regions in DNA usually begins with the analysis of nested sets of deletions that
penetrate into the region of interest from the upstream and downstream directions
.
Linker - scanning Mutagenesis The identification and localization of regulatory
regions in DNA usually begins with the analysis of nested sets of deletions that
penetrate into the region of interest from the upstream and downstream directions
.
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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