Molecular Cloning: A Laboratory Manual, Book 2 |
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Page 5
The first step in both procedures is the generation of a set of circularly permuted
linear molecules by random double - stranded cleavage of superhelical plasmid
DNA . The two methods differ in the techniques used to achieve this random ...
The first step in both procedures is the generation of a set of circularly permuted
linear molecules by random double - stranded cleavage of superhelical plasmid
DNA . The two methods differ in the techniques used to achieve this random ...
Page 9
While the tubes are incubating , digest approximately 1 ug of the plasmid DNA
with an excess ( 5 units ) of a restriction enzyme that cleaves the DNA only once .
The resulting linear DNA will be used as an electrophoretic marker ( see h ) . g .
While the tubes are incubating , digest approximately 1 ug of the plasmid DNA
with an excess ( 5 units ) of a restriction enzyme that cleaves the DNA only once .
The resulting linear DNA will be used as an electrophoretic marker ( see h ) . g .
Page 89
Under these conditions of electrophoresis , closed circular DNA migrates slightly
faster than linear DNA and considerably faster than relaxed circular DNA . d .
Examine the gel by ultraviolet illumination , and determine the conditions that
give ...
Under these conditions of electrophoresis , closed circular DNA migrates slightly
faster than linear DNA and considerably faster than relaxed circular DNA . d .
Examine the gel by ultraviolet illumination , and determine the conditions that
give ...
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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