Molecular Cloning: A Laboratory Manual, Book 2 |
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Page 12-38
... 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 × 108 pfu of the bac- teriophage stock . The ...
... 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 × 108 pfu of the bac- teriophage stock . The ...
Page 12-39
... cultures to a water bath set at 44 ° C than to a hot - air incubator . Shake the cultures vigorously from time to time during incubation at 44 ° C . 11. Add isopropylthio - B - D - galactoside ( IPTG ) to each culture to a final con ...
... cultures to a water bath set at 44 ° C than to a hot - air incubator . Shake the cultures vigorously from time to time during incubation at 44 ° C . 11. Add isopropylthio - B - D - galactoside ( IPTG ) to each culture to a final con ...
Page 15-76
... culture of E. coli strain CJ236 ( dut ̄ ung ̄ F ' ) ( Kunkel et al . 1987 ) . Incubate the culture with vigorous shaking ( 300 cycles / minute on a rotary shaker ) for 6 hours at 37 ° C . The bacteriophage suspension used as an inoculum ...
... culture of E. coli strain CJ236 ( dut ̄ ung ̄ F ' ) ( Kunkel et al . 1987 ) . Incubate the culture with vigorous shaking ( 300 cycles / minute on a rotary shaker ) for 6 hours at 37 ° C . The bacteriophage suspension used as an inoculum ...
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