Molecular Cloning: A Laboratory Manual, Book 3 |
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Page 32
Centrifuge at 12 , 000g for 1 minute at 4°C in a microfuge to pellet the cells .
Discard the supernatant . b . Suspend the cell pellets in 100 ul of a freshly
prepared solution of lysozyme ( 1 mg / ml ) , 20 % w / v sucrose , 30 mm Tris · Cl (
pH 8 .
Centrifuge at 12 , 000g for 1 minute at 4°C in a microfuge to pellet the cells .
Discard the supernatant . b . Suspend the cell pellets in 100 ul of a freshly
prepared solution of lysozyme ( 1 mg / ml ) , 20 % w / v sucrose , 30 mm Tris · Cl (
pH 8 .
Page 40
1 . Centrifuge the cell lysate at 12 , 000g for 15 minutes at 4°C in a microfuge . 2 .
Decant the supernatant . Resuspend the pellet in 1 ml of H , O per gram of E . coli
. Transfer 100 - ul aliquots to four microfuge tubes and store the remainder . 3 .
1 . Centrifuge the cell lysate at 12 , 000g for 15 minutes at 4°C in a microfuge . 2 .
Decant the supernatant . Resuspend the pellet in 1 ml of H , O per gram of E . coli
. Transfer 100 - ul aliquots to four microfuge tubes and store the remainder . 3 .
Page 40
After induction for the appropriate period of time , recover the bacteria from 1 ml
of culture by centrifugation at 12 , 000g for 30 seconds in a microfuge . 2 .
Remove the medium by aspiration , and then resuspend the pellet by vortexing in
0 .
After induction for the appropriate period of time , recover the bacteria from 1 ml
of culture by centrifugation at 12 , 000g for 30 seconds in a microfuge . 2 .
Remove the medium by aspiration , and then resuspend the pellet by vortexing in
0 .
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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
Cosmid Vectors | 3 |
Introduction of Recombinant Vectors into Mammalian Cells 16 | 30 |
TRANSFECTION MEDIATED BY DEAEDEXTRAN 16 | 41 |
Copyright | |
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Common terms and phrases
acetate acid activity added Adjust aliquots allow amount antibody antigen Appendix appropriate approximately assay bacteria bacteriophage binding buffer carried cells centrifugation Cl pH cloned coli column complete concentration containing culture deionized water described desired detected determined dilution Dissolve efficiency electrophoresis enzyme et al ethanol expression extract Figure filter final foreign fragment gene growth increase Incubate levels lines mammalian cells medium method mg/ml microfuge minutes mixture mRNA nucleic acid obtained pellet Place plasmid plate polymerase precipitate prepared presence production promoter protein radioactive radiolabeled reaction Remove restriction room temperature sample selection sequences sodium solution specific step sterile stored strain supernatant TABLE target protein Telephone termini tion transcription transfected Transfer transformation translation Tris tube ug/ml usually vectors volume wash western blotting ОН
Bibliographic information
| Title | Molecular Cloning: A Laboratory Manual, Book 3 Molecular Cloning: A Laboratory Manual, Joseph Sambrook, ISBN 0879693096, 9780879693091 |
| Authors | Joseph Sambrook, E. F. Fritsch, Tom Maniatis |
| Edition | 2 |
| Publisher | Cold Spring Harbor Laboratory, 1989 |
| Original from | the University of Michigan |
| Digitized | 23 Feb 2010 |
| ISBN | 0879693096, 9780879693091 |
| Length | 412 pages |
| Export Citation | BiBTeX EndNote RefMan |