Molecular Cloning: A Laboratory Manual, Book 2 |
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Page 4
In general , such oligonucleotides match with their target sequence perfectly , or
nearly perfectly , and they are sufficiently long ( 19 – 40 nucleotides ) to allow the
use of hybridization conditions that can guarantee discrimination between the ...
In general , such oligonucleotides match with their target sequence perfectly , or
nearly perfectly , and they are sufficiently long ( 19 – 40 nucleotides ) to allow the
use of hybridization conditions that can guarantee discrimination between the ...
Page 55
EMPIRICAL DETERMINATION OF MELTING TEMPERATURE The melting
temperature ( Tm ) of an oligonucleotide hybridized to a target sequence can be
determined by the procedure described below . The protocol actually measures
the ...
EMPIRICAL DETERMINATION OF MELTING TEMPERATURE The melting
temperature ( Tm ) of an oligonucleotide hybridized to a target sequence can be
determined by the procedure described below . The protocol actually measures
the ...
Page 32
For each amplification reaction , measure the ratio of the yield of amplified control
template to the yield of amplified target sequence . Plot this ratio against the
amount of control template added to each amplification reaction . From the
resulting ...
For each amplification reaction , measure the ratio of the yield of amplified control
template to the yield of amplified target sequence . Plot this ratio against the
amount of control template added to each amplification reaction . From the
resulting ...
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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