Molecular Cloning: A Laboratory Manual, Book 2 |
From inside the book
Results 1-3 of 83
Page 13-82
... appropriate [ 32P ] dNTP and the Klenow fragment of E. coli DNA polymerase I. In the example shown in Figure 13.11 , a recombinant plasmid carrying the target at the central SmaI site is cleaved with Tth1111 , generating two fragments ...
... appropriate [ 32P ] dNTP and the Klenow fragment of E. coli DNA polymerase I. In the example shown in Figure 13.11 , a recombinant plasmid carrying the target at the central SmaI site is cleaved with Tth1111 , generating two fragments ...
Page 13-86
... appropriate waste bottle . A + G Mix 10 μl H2O 4 μl sonicated DNA 10 μl 32P - labeled DNA Chill to 0 ° C . Add 4 μl piperidine for- mate ( pH 2.0 ) and mix . Incubate 15 minutes at 37 ° C . Add 240 μl hydrazine stop solution ( at 0 ° C ) ...
... appropriate waste bottle . A + G Mix 10 μl H2O 4 μl sonicated DNA 10 μl 32P - labeled DNA Chill to 0 ° C . Add 4 μl piperidine for- mate ( pH 2.0 ) and mix . Incubate 15 minutes at 37 ° C . Add 240 μl hydrazine stop solution ( at 0 ° C ) ...
Page 15-30
... appropriate 10 × restriction enzyme buffer the appropriate restriction enzyme 22 μl 5 με 5-10 units Incubate the reaction for 2 hours at the appropriate temperature . 16. Transform ( plasmids or phagemids ) or transfect ( bacteriophage ...
... appropriate 10 × restriction enzyme buffer the appropriate restriction enzyme 22 μl 5 με 5-10 units Incubate the reaction for 2 hours at the appropriate temperature . 16. Transform ( plasmids or phagemids ) or transfect ( bacteriophage ...
Other editions - View all
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