Molecular Cloning: A Laboratory Manual, Volume 1 |
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Page 190
SCREENING BY HYBRIDIZATION In 1975 , Grunstein and Hogness described a
method for in situ lysis of bacterial colonies on nitrocellulose filters and ... The
DNA could then be hybridized to appropriate radiolabeled nucleic acid probes .
SCREENING BY HYBRIDIZATION In 1975 , Grunstein and Hogness described a
method for in situ lysis of bacterial colonies on nitrocellulose filters and ... The
DNA could then be hybridized to appropriate radiolabeled nucleic acid probes .
Page 7-39
NORTHERN HYBRIDIZATION Initially , northern hybridization was carried out
exclusively with RNA immobilized on diazotized cellulose (
diazobenzyloxymethyl ( DBM ) -cellulose ) ( Alwine et al . 1977 ) ( see Chapter 9 ,
pages 9.35–9.36 ) .
NORTHERN HYBRIDIZATION Initially , northern hybridization was carried out
exclusively with RNA immobilized on diazotized cellulose (
diazobenzyloxymethyl ( DBM ) -cellulose ) ( Alwine et al . 1977 ) ( see Chapter 9 ,
pages 9.35–9.36 ) .
Page 7-75
It is often difficult to obtain complete dissolution in hybridization buffer of RNA that
has been precipitated with ethanol . The problem is exacerbated if the pellet is
dried in a desiccator . Sometimes the RNA can be redissolved by a combination ...
It is often difficult to obtain complete dissolution in hybridization buffer of RNA that
has been precipitated with ethanol . The problem is exacerbated if the pellet is
dried in a desiccator . Sometimes the RNA can be redissolved by a combination ...
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Common terms and phrases
activity agar agarose aliquots allow amount appropriate arms bacterial bacteriophage BamHI buffer carrying cells centrifugation Chapter cloning coli colonies completely concentration construction containing cosmid culture described digestion EcoRI efficiency electrophoresis et al ethanol ethidium ethidium bromide extracts Figure filters final foreign DNA fragments gene genetic genome growth Hindi host hybridization Incubate infected inserted libraries ligase ligation linear method microfuge minutes minutes at 4°C mixture molecules mutations nucleic acids obtained original packaging particles pellet plaques plasmid DNA plate polycloning polymerase possible precipitate prepared presence probes problem procedure promoter protein purified reaction recombinant Recover region remove replication restriction enzyme room temperature Sall segment selection sequences single single-stranded Smal solution step sterile stored strains strand supernatant termini transfer transformation tube usually vector volume Xbal yield