Molecular Cloning: A Laboratory Manual, Volume 1 |
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Page 1-48
... ethanol to drain away . 7. Wash the pellet with 70 % ethanol at 4 ° C , and recentrifuge as in step 5 . Remove the supernatant as completely as possible , and store the open tube for a few minutes in an inverted position on a pad of ...
... ethanol to drain away . 7. Wash the pellet with 70 % ethanol at 4 ° C , and recentrifuge as in step 5 . Remove the supernatant as completely as possible , and store the open tube for a few minutes in an inverted position on a pad of ...
Page 7-14
... ethanol to evaporate . 18. Redissolve the pellet in 200 μl of TE ( pH 7.6 ) . Add 500 μl of ethanol , and store the preparation at -70 ° C until it is needed . To recover the RNA , remove an aliquot , add 3 м sodium acetate ( pH 5.2 ) ...
... ethanol to evaporate . 18. Redissolve the pellet in 200 μl of TE ( pH 7.6 ) . Add 500 μl of ethanol , and store the preparation at -70 ° C until it is needed . To recover the RNA , remove an aliquot , add 3 м sodium acetate ( pH 5.2 ) ...
Page 7-21
... ethanol at room temperature . Invert the tube and drain off the ethanol , again checking that the pellet of RNA is not dislodged . Washing with ethanol removes CsCl from the pellet of RNA , making it easier to dissolve . 9. Allow the ...
... ethanol at room temperature . Invert the tube and drain off the ethanol , again checking that the pellet of RNA is not dislodged . Washing with ethanol removes CsCl from the pellet of RNA , making it easier to dissolve . 9. Allow the ...
Contents
DEVELOPMENT OF PLASMID CLONING VECTORS | 1-7 |
Constructing Expression Libraries in Plasmid and Bacteriophage | 1-12 |
Expression Libraries Constructed in Bacteriophage 12 | 1-19 |
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agar plate agarose gel aliquots amber mutations ampicillin antibiotic bacteriophage particles bacteriophage T4 bacteriophage T4 DNA BamHI Bgill buffer carrying cDNA cells cloning coli cosmid cosmid vector culture digestion DNA fragments DNA ligase DNA molecules DNA polymerase double-stranded DNA EcoRI EDTA EDTA pH 8.0 efficiency Escherichia coli ethanol ethidium bromide eukaryotic eukaryotic DNA exonuclease extracts foreign DNA formamide gene genetic HindIII Hindill host hours at 37°C hybridization Incubate infected inserted Kpnl lacZ ligation reaction linear lysogenic method microfuge tube minutes at 4°C nin5 nitrocellulose nitrocellulose filter Nucleic Acids nucleotides packaging pasteur pipette pellet plaques plasmid DNA plasmid vectors polycloning prepared probes protein purified Pvul recA red gam remove replication restriction enzyme RNAase room temperature Sacl Sall sequences Smal solution sterile stored strains strand stuffer supernatant T4 DNA ligase teriophage transfer transformation Tris Cl pH vector DNA vitro volume Xbal µg/ml