Molecular Cloning: A Laboratory Manual, Book 2 |
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Page 9-16
... centrifuge tube . Recover the cells by centrifugation at 1500g for 10 minutes at 4 ° C . Resuspend the cells in 5-10 volumes of ice - cold TBS and repeat the centrifugation . Resuspend the cells in TE ( pH 8.0 ) at a concentration of 5 ...
... centrifuge tube . Recover the cells by centrifugation at 1500g for 10 minutes at 4 ° C . Resuspend the cells in 5-10 volumes of ice - cold TBS and repeat the centrifugation . Resuspend the cells in TE ( pH 8.0 ) at a concentration of 5 ...
Page 14-23
... centrifuge , and take off the retentate cup . Add 0.75 ml of H2O to the sample reservoir , replace the retentate cup , and recentrifuge as described in step 4 . 6. Repeat step 5 twice more . At the end of the fourth centrifugation , the ...
... centrifuge , and take off the retentate cup . Add 0.75 ml of H2O to the sample reservoir , replace the retentate cup , and recentrifuge as described in step 4 . 6. Repeat step 5 twice more . At the end of the fourth centrifugation , the ...
Page 15-77
... centrifugation at 4000g for 5 minutes at room temperature . Avoid transferring material from the interface . 11. Transfer the aqueous phase from the final extraction to a glass centrifuge tube ( e.g. , a 30 - ml Corex tube ) . Measure ...
... centrifugation at 4000g for 5 minutes at room temperature . Avoid transferring material from the interface . 11. Transfer the aqueous phase from the final extraction to a glass centrifuge tube ( e.g. , a 30 - ml Corex tube ) . Measure ...
Contents
E | 8-8 |
SYNTHESIS OF THE FIRST STRAND OF CDNA 8 | 8-11 |
CONCENTRATING NUCLEIC ACIDS BY EXTRACTION WITH BUTANOL E | 8-16 |
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acrylamide agarose gel aliquots amplification annealing antibody bacteriophage M13 bacteriophage T4 bacteriophage T4 DNA buffer carried cDNA clones cDNA libraries cells centrifugation at 12,000g cleavage cleaved cleotide coli DNA polymerase concentration containing ddNTPs deletions denatured digestion diluted dithiothreitol DNA ligase DNA sequencing DNAase dNTPs double-stranded DNA EDTA EDTA pH 8.0 ethanol exonuclease expression libraries formamide fusion proteins gel electrophoresis gene genomic DNA H₂O hybridization Klenow fragment labeled ligation linear linkers Maxam-Gilbert method MgCl2 microfuge tube minutes at 4°C molecules mRNA Natl nitrocellulose nitrocellulose filters Nucleic Acids nucleotides oligonucleotide phagemid piperidine plaques plasmid plasmid DNA plate polymerase chain reaction precipitation prepared primer probe purified radioactivity radiolabeled recombinant remove restriction enzyme room temperature samples Sequenase sequencing gel sequencing reactions sonicated specific activity stored strand of cDNA synthesis T4 DNA polymerase target DNA target sequence template DNA termini transcription transfer Tris Cl pH vectors µg/ml