Molecular Cloning: A Laboratory Manual, Book 2 |
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Page 10-52
... appropriate restriction enzyme buffer . 2. Add 2-10 μCi ( sp . act . 400-3000 Ci / mmole ) of the appropriate [ a- 32P ] dNTP . 3. Add 1 unit of the Klenow fragment of E. coli DNA polymerase I , and incubate the reaction for 15 minutes ...
... appropriate restriction enzyme buffer . 2. Add 2-10 μCi ( sp . act . 400-3000 Ci / mmole ) of the appropriate [ a- 32P ] dNTP . 3. Add 1 unit of the Klenow fragment of E. coli DNA polymerase I , and incubate the reaction for 15 minutes ...
Page 15-25
... appropriate 10 × restriction enzyme buffer and 8 units of a restriction enzyme that will separate the target DNA from the vector . Incubate the reactions for 1 hour at the appropriate temperature . 13. At the end of the incubation ...
... appropriate 10 × restriction enzyme buffer and 8 units of a restriction enzyme that will separate the target DNA from the vector . Incubate the reactions for 1 hour at the appropriate temperature . 13. At the end of the incubation ...
Page 15-30
... appropriate 10 x 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 x 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 ...
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