Molecular Cloning: A Laboratory Manual, Volume 1 |
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Page 154
The fragment of foreign DNA is then inserted into the vector by a process known
as directional cloning ( Figure 1.6 ) . For example , the vector pUC19 can be
digested with BamHI and HindiII and , after digestion , the large fragment of the
vector ...
The fragment of foreign DNA is then inserted into the vector by a process known
as directional cloning ( Figure 1.6 ) . For example , the vector pUC19 can be
digested with BamHI and HindiII and , after digestion , the large fragment of the
vector ...
Page 187
The DNA to be inserted and the purified plasmid DNA are digested with
restriction enzymes that recognize sites located in only one of the antibiotic
resistance genes ( in this example , the tet " gene ) . After ligating the two DNAs at
the ...
The DNA to be inserted and the purified plasmid DNA are digested with
restriction enzymes that recognize sites located in only one of the antibiotic
resistance genes ( in this example , the tet " gene ) . After ligating the two DNAs at
the ...
Page 3-19
pCV103 , pCV107 , pCV108 These are derivatives of pJB8 ( amp " , ColEl origin )
that carry selectable markers for eukaryotic cells on segments of DNA inserted at
the Ball site of PJB8 ( Lau and Kan 1983 ) . PCV103 contains the E. coli ...
pCV103 , pCV107 , pCV108 These are derivatives of pJB8 ( amp " , ColEl origin )
that carry selectable markers for eukaryotic cells on segments of DNA inserted at
the Ball site of PJB8 ( Lau and Kan 1983 ) . PCV103 contains the E. coli ...
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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