Molecular Cloning: A Laboratory Manual, Book 2 |
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Page 13-21
... target DNAs into segments that are optimal in size for random sequencing ... DNA in a random fashion that is relatively independent of sequence , and ... DNA is digested to completion with a single restriction enzyme . In theory , sets of ...
... target DNAs into segments that are optimal in size for random sequencing ... DNA in a random fashion that is relatively independent of sequence , and ... DNA is digested to completion with a single restriction enzyme . In theory , sets of ...
Page 13-24
... Target DNA 1. Using the appropriate restriction enzymes , digest a sufficient amount of recombinant vector to yield 10-15 μg of purified target DNA . Separate the target DNA from the vector by preparative gel electrophoresis , and recover ...
... Target DNA 1. Using the appropriate restriction enzymes , digest a sufficient amount of recombinant vector to yield 10-15 μg of purified target DNA . Separate the target DNA from the vector by preparative gel electrophoresis , and recover ...
Page 15-48
A Laboratory Manual Joseph Sambrook, Tom Maniatis. EXCISION OF THE TARGET DNA 1. Excise the target DNA by digesting approximately 50 μg of the recircular- ized plasmid DNA from step 6 , page 15.47 , with the appropriate restric- tion ...
A Laboratory Manual Joseph Sambrook, Tom Maniatis. EXCISION OF THE TARGET DNA 1. Excise the target DNA by digesting approximately 50 μg of the recircular- ized plasmid DNA from step 6 , page 15.47 , with the appropriate restric- tion ...
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Common terms and phrases
agarose gel aliquots amino acid amplification bacteriophage M13 bacteriophage T4 bacteriophage T4 DNA Biol buffer carried cDNA libraries cells centrifugation at 12,000g Chapter cleavage cleaved cleotide codon coli DNA polymerase complementary concentration containing deletions denatured digestion DNA fragments DNA ligase DNA sequencing DNAase dNTPs double-stranded DNA EDTA EDTA pH 8.0 efficiency ethanol ethidium bromide exonuclease formamide gel electrophoresis gene genomic DNA H₂O hybridization Incubate the reaction inserted Klenow fragment labeled ligation linkers method microfuge tube minutes at 4°C mismatched molecules mRNA mutagenesis mutagenic oligonucleotide mutations Natl nitrocellulose nucleotides oligonu oligonucleotide phagemid plaques plasmid plasmid DNA plates polymerase chain reaction pool primer probe protein purified radioactivity radiolabeled recombinant remove restriction enzyme room temperature samples screening sequencing reactions single-stranded DNA solution specific activity Store strand of cDNA synthesis target DNA target fragment target sequence template DNA termini transcription transfer Tris Cl pH vector µg/ml