Molecular Cloning: A Laboratory Manual, Book 2 |
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Page 11-21
Preparation of Synthetic Oligonucleotides Oligonucleotides are usually supplied
as a lyophilized powder . 1 . In a sterile microfuge tube , dissolve the crude
oligonucleotide ( usually 1 - 10 nmoles ) in 1 ml of sterile , filtered H , O ( Milli - Q
or ...
Preparation of Synthetic Oligonucleotides Oligonucleotides are usually supplied
as a lyophilized powder . 1 . In a sterile microfuge tube , dissolve the crude
oligonucleotide ( usually 1 - 10 nmoles ) in 1 ml of sterile , filtered H , O ( Milli - Q
or ...
Page 11-30
Elute the oligonucleotide three times with 1 ml of a 60 : 40 mixture of methanol : H
, O . Repeat step lc after each elution . Collect each effluent in a separate
microfuge tube . Read the OD260 of the solution in each of the three microfuge
tubes ...
Elute the oligonucleotide three times with 1 ml of a 60 : 40 mixture of methanol : H
, O . Repeat step lc after each elution . Collect each effluent in a separate
microfuge tube . Read the OD260 of the solution in each of the three microfuge
tubes ...
Page 11-31
LABELING OF SYNTHETIC OLIGONUCLEOTIDES BY PHOSPHORYLATION
WITH BACTERIOPHAGE T4 POLYNUCLEOTIDE ... The reaction described
below is designed to label 10 pmoles of an oligonucleotide to high specific
activity .
LABELING OF SYNTHETIC OLIGONUCLEOTIDES BY PHOSPHORYLATION
WITH BACTERIOPHAGE T4 POLYNUCLEOTIDE ... The reaction described
below is designed to label 10 pmoles of an oligonucleotide to high specific
activity .
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LibraryThing Review
User Review - iayork - LibraryThingthe BIBLE of every biologist: So few and so much to say about this bible of Biology at the bench... You'll really find everything you want in it, including the composition of all the buffers and ... Read full review
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Common terms and phrases
activity addition aliquots allow amount amplification appropriate approximately bacteriophage bacteriophage T4 base buffer carried cDNA cells centrifugation Chapter chromatography cleavage cleaved clones complete concentration constructed containing deletions denatured described determine digestion dNTPs double-stranded efficiency electrophoresis et al ethanol expression extraction Figure filter four fragment gene genomic DNA hybridization increase Incubate inserted interest isolation Klenow labeled length libraries ligation linear linkers membrane method minutes mixture molecules mRNA mutations nucleic acids nucleotides obtained oligonucleotide original 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 specific step stored strand synthesis target DNA target sequence template termini transfer Tris tube units usually vector volume