Molecular Cloning: A Laboratory Manual, Book 2 |
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Page 8-65
... nuclease - S1 buffer . 10x Nuclease - S1 buffer 2 M NaCl 0.5 M sodium acetate ( pH 4.5 ) 10 mM ZnSO 5 % glycerol f . Add 5 units of nuclease S1 to one of the two tubes containing the first strand of cDNA and to one containing the second ...
... nuclease - S1 buffer . 10x Nuclease - S1 buffer 2 M NaCl 0.5 M sodium acetate ( pH 4.5 ) 10 mM ZnSO 5 % glycerol f . Add 5 units of nuclease S1 to one of the two tubes containing the first strand of cDNA and to one containing the second ...
Page 15-39
... nuclease S1 ) . Superhelical DNA is under torsional strain and may therefore be susceptible to cleavage at particular sites by nuclease S1 . Digestion by nuclease S1 at these sites would generate linear molecules whose termini would map ...
... nuclease S1 ) . Superhelical DNA is under torsional strain and may therefore be susceptible to cleavage at particular sites by nuclease S1 . Digestion by nuclease S1 at these sites would generate linear molecules whose termini would map ...
Page 15-40
... nuclease S1 1. Digest the nicked / gapped circular DNA with nuclease S1 as follows : a . Redissolve the DNA purified from the CsCl - ethidium bromide gradient ( 60-100 μg ) ( step 7 , page 15.39 ) in 500 μl of 1 × nuclease - S1 buffer ...
... nuclease S1 1. Digest the nicked / gapped circular DNA with nuclease S1 as follows : a . Redissolve the DNA purified from the CsCl - ethidium bromide gradient ( 60-100 μg ) ( step 7 , page 15.39 ) in 500 μl of 1 × nuclease - S1 buffer ...
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