Molecular Cloning: A Laboratory Manual, Book 2 |
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Page 11-34
Using an automatic pipetting device equipped with a disposable tip , carefully
remove all of the supernatant fluid ( which contains most of the unincorporated [ y
- " ? P ] ATP and any free 32P generated during the phosphorylation reaction ) . 3
.
Using an automatic pipetting device equipped with a disposable tip , carefully
remove all of the supernatant fluid ( which contains most of the unincorporated [ y
- " ? P ] ATP and any free 32P generated during the phosphorylation reaction ) . 3
.
Page 12-27
Adjust the pH of the supernatant to 9 . 0 with 1 n NaOH . 10 . Chill the extract to 0°
C , and then bind the bacterial proteins to cyanogenbromide - activated
Sepharose 4B ( Pharmacia ) according to the manufacturer ' s instructions . 11 .
Before ...
Adjust the pH of the supernatant to 9 . 0 with 1 n NaOH . 10 . Chill the extract to 0°
C , and then bind the bacterial proteins to cyanogenbromide - activated
Sepharose 4B ( Pharmacia ) according to the manufacturer ' s instructions . 11 .
Before ...
Page 15-76
It is important to kill the bacterial cells to prevent them from continuing to produce
thymine - containing viral DNA during the next round of bacteriophage growth . 3
. Transfer 50 ul of the supernatant to a 500 - ml flask containing 50 ml of 2 x YT ...
It is important to kill the bacterial cells to prevent them from continuing to produce
thymine - containing viral DNA during the next round of bacteriophage growth . 3
. Transfer 50 ul of the supernatant to a 500 - ml flask containing 50 ml of 2 x YT ...
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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