Molecular Cloning: A Laboratory Manual, Book 2 |
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Page 8-24
It is therefore important to arrange the ligation conditions to minimize the
formation of these chimeric molecules , since it is extremely laborious to test
subsequently whether cDNA clones that contain the particular restriction site at
an internal ...
It is therefore important to arrange the ligation conditions to minimize the
formation of these chimeric molecules , since it is extremely laborious to test
subsequently whether cDNA clones that contain the particular restriction site at
an internal ...
Page 13-24
GENERATION OF A LIBRARY OF RANDOMLY OVERLAPPING CLONES
Purification and Ligation of the Target DNA 1 . ... Because the next step of the
procedure requires the target DNA to be ligated to itself , it is best to use
restriction ...
GENERATION OF A LIBRARY OF RANDOMLY OVERLAPPING CLONES
Purification and Ligation of the Target DNA 1 . ... Because the next step of the
procedure requires the target DNA to be ligated to itself , it is best to use
restriction ...
Page 15-10
Add 1 - 2 Weiss units of bacteriophage T4 DNA ligase , and incubate the reaction
for 6 - 8 hours at 16°C . 10 x Blunt - end ... Blunt - end ligation buffer should be
stored in small aliquots at - 20°C . The spermidine and hexamminecobalt
chloride ...
Add 1 - 2 Weiss units of bacteriophage T4 DNA ligase , and incubate the reaction
for 6 - 8 hours at 16°C . 10 x Blunt - end ... Blunt - end ligation buffer should be
stored in small aliquots at - 20°C . The spermidine and hexamminecobalt
chloride ...
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LibraryThing Review
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Contents
CONCENTRATING NUCLEIC ACIDS E | 8-10 |
SYNTHESIS OF THE FIRST STRAND OF DNA 8 | 8-11 |
MEASUREMENT OF RADIOACTIVITY IN NUCLEIC ACIDS E | 8-18 |
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Common terms and phrases
aliquots allow amount amplification antibody appropriate approximately bacteriophage bacteriophage T4 base binding buffer carried cDNA cells centrifugation Chapter chromatography cleaved clones colonies complete concentration constructed containing deletions denatured described detection determine digestion dNTPs double-stranded efficiency electrophoresis et al ethanol expression extraction Figure filters four fragment gene genomic DNA hybridization increase Incubate inserted interest isolated Klenow labeled length libraries ligation linear linkers membrane method minutes mixture molecules mRNA mutants nucleic acids nucleotides obtained oligonucleotide plaques plasmid DNA plate position possible precipitation prepared presence primer probe problem protein purified radioactivity radiolabeled reaction recombinant region remove restriction enzyme room temperature samples screening separate sequence single single-stranded solution step stored strand synthesis target DNA target sequence template termini transfer Tris tube units usually vector volume