Molecular Cloning: A Laboratory Manual, Book 2 |
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Page 13-24
... step 2. If it is not possible to avoid restriction enzymes that generate blunt ends , the ligation ( step 2 ) should be carried out in the presence of substances that increase macromolecular crowding ( e.g. , polyethylene glycol ...
... step 2. If it is not possible to avoid restriction enzymes that generate blunt ends , the ligation ( step 2 ) should be carried out in the presence of substances that increase macromolecular crowding ( e.g. , polyethylene glycol ...
Page 14-32
... ( step 2 ) . Do not reduce the concentration of unlabeled dCTP in the reaction mixture to increase the specific activity of the precursor pool . There is a danger that the concentration of the nucleotide could become limiting at late ...
... ( step 2 ) . Do not reduce the concentration of unlabeled dCTP in the reaction mixture to increase the specific activity of the precursor pool . There is a danger that the concentration of the nucleotide could become limiting at late ...
Page 15-46
... step is no longer optimal . 12. Just before plating on selective media , pool the several small - scale transformation reactions and recover the bacterial cells by centrifugation at 5000g for 10 minutes at room temperature . Resuspend ...
... step is no longer optimal . 12. Just before plating on selective media , pool the several small - scale transformation reactions and recover the bacterial cells by centrifugation at 5000g for 10 minutes at room temperature . Resuspend ...
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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