Molecular Cloning: A Laboratory Manual, Book 2 |
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Results 1-3 of 45
Page 8-77
... ligations as a guide , set up a large - scale ligation reaction , increasing the amounts of all components proportionately . Be sure to set up a control reaction that contains only bacteriophage λ arms and no cDNA . Package all of the ...
... ligations as a guide , set up a large - scale ligation reaction , increasing the amounts of all components proportionately . Be sure to set up a control reaction that contains only bacteriophage λ arms and no cDNA . Package all of the ...
Page 13-31
... ligation reactions are to work efficiently . An alternative method is to heat the reaction ( at the end of step 2 ) to 65 ° C for 1 hour ( or 75 ° C for 10 minutes ) and then to extract once with phenol : chloroform . 4. Extract the ...
... ligation reactions are to work efficiently . An alternative method is to heat the reaction ( at the end of step 2 ) to 65 ° C for 1 hour ( or 75 ° C for 10 minutes ) and then to extract once with phenol : chloroform . 4. Extract the ...
Page 15-30
... ligation reaction should not exceed 5-6 nM . This concentration is equivalent to 10 μg / ml of a plasmid of the size of pBR322 . If desired , a phosphorylated linker can be inserted at the site of recircularization by including the ...
... ligation reaction should not exceed 5-6 nM . This concentration is equivalent to 10 μg / ml of a plasmid of the size of pBR322 . If desired , a phosphorylated linker can be inserted at the site of recircularization by including the ...
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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