Molecular Cloning: A Laboratory Manual, Book 2 |
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Page 10-64
... for 10 minutes at 4 ° C in a microfuge . • 2. Dissolve the DNA in a minimum volume of 10 mM Tris Cl ( pH 8.3 ) . 3. Add : 10 CIP dephosphorylation buffer H2O to 48 μl 10x CIP dephosphorylation buffer 10 mм ZnCl2 10 mM MgCl2 100 mM Tris ...
... for 10 minutes at 4 ° C in a microfuge . • 2. Dissolve the DNA in a minimum volume of 10 mM Tris Cl ( pH 8.3 ) . 3. Add : 10 CIP dephosphorylation buffer H2O to 48 μl 10x CIP dephosphorylation buffer 10 mм ZnCl2 10 mM MgCl2 100 mM Tris ...
Page 12-18
... 4 ° C until the results of the immunological screening are available . If large areas of the top agarose stick to the nitrocellulose filters , chill the plates for 30 minutes at 4 ° C or 5 minutes at -20 ° C before peeling off the ...
... 4 ° C until the results of the immunological screening are available . If large areas of the top agarose stick to the nitrocellulose filters , chill the plates for 30 minutes at 4 ° C or 5 minutes at -20 ° C before peeling off the ...
Page 13-96
... for 30 seconds . A > C Mix 4 μl sonicated DNA 5 μl 32P - labeled DNA Add 1 μl 30 % NaOH . Incubate 6 minutes at 90 ° C . Chill to 0 ° C . Add 150 μl 1M piperidine ( at 0 ° C ) . Store on ice until all reactions are ready for incubation .
... for 30 seconds . A > C Mix 4 μl sonicated DNA 5 μl 32P - labeled DNA Add 1 μl 30 % NaOH . Incubate 6 minutes at 90 ° C . Chill to 0 ° C . Add 150 μl 1M piperidine ( at 0 ° C ) . Store on ice until all reactions are ready for incubation .
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Common terms and phrases
agarose gel aliquots amplification annealing antibody bacterial bacteriophage M13 bacteriophage T4 bacteriophage T4 DNA buffer carried cDNA clones cDNA libraries cells centrifugation at 12,000g Chapter cleavage cleaved cleotide codon coli DNA polymerase concentration containing deletions denatured digestion dithiothreitol DNA ligase DNA sequencing DNAase dNTPs double-stranded DNA EDTA EDTA pH 8.0 efficiency ethanol exonuclease formamide fusion proteins gel electrophoresis gene genomic DNA H₂O hybridization Incubate inserted Klenow fragment ligation linear linkers method microfuge tube minutes at 4°C molecules mRNA mutagenesis mutagenic oligonucleotide mutations Natl nitrocellulose nitrocellulose filters Nucleic Acids nucleotides oligonucleotide phagemid plaques plasmid plasmid DNA plate polymerase chain reaction prepared primer probes purified radioactivity radiolabeled reaction mixture recombinant remove restriction enzyme reverse transcriptase room temperature samples screening sequencing reactions single-stranded DNA solution step Store strand of cDNA synthesis target DNA target sequence template DNA termini transfer Tris Cl pH vector