Molecular Cloning: A Laboratory Manual, Book 2 |
From inside the book
Results 1-3 of 85
Page 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 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 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 .
What people are saying - Write a review
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
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
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 |
Copyright | |
92 other sections not shown
Other editions - View all
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