Molecular Cloning: A Laboratory Manual, Book 2 |
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Page 12-38
... MgCl2 . 3. Dilute 50 μl of the saturated culture with 2 ml of LB medium containing 10 mM MgCl2 . Transfer four 100 - μl aliquots of the diluted culture to fresh culture tubes . 4. To three of the tubes , add 1 × 107 , 5 × 107 , or 2 ...
... MgCl2 . 3. Dilute 50 μl of the saturated culture with 2 ml of LB medium containing 10 mM MgCl2 . Transfer four 100 - μl aliquots of the diluted culture to fresh culture tubes . 4. To three of the tubes , add 1 × 107 , 5 × 107 , or 2 ...
Page 13-41
... MgCl2 100 mм dithiothreitol 500 μg / ml bovine serum albumin ( Fraction V ; Sigma ) ( optional ) This buffer should be stored in small aliquots at -20 ° C . For definition of Weiss unit , see page 13.25 . 11. Transform ( phagemid ...
... MgCl2 100 mм dithiothreitol 500 μg / ml bovine serum albumin ( Fraction V ; Sigma ) ( optional ) This buffer should be stored in small aliquots at -20 ° C . For definition of Weiss unit , see page 13.25 . 11. Transform ( phagemid ...
Page 15-29
... MgCl2 1 mм dithiothreitol The use of two different DNA polymerases in the repair reaction results in an approximately threefold increase in recovery of mutants . 11. Add 70 μl of TE ( pH 7.6 ) , and then extract the sample once with an ...
... MgCl2 1 mм dithiothreitol The use of two different DNA polymerases in the repair reaction results in an approximately threefold increase in recovery of mutants . 11. Add 70 μl of TE ( pH 7.6 ) , and then extract the sample once with an ...
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