Molecular Cloning: A Laboratory Manual, Book 2 |
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Page 11-21
... sterile microfuge tube , dissolve the crude oligonucleotide ( usually 1-10 nmoles ) in 1 ml of sterile , filtered H2O ( Milli - Q or equivalent ) . Vortex the solution thoroughly . The solution is often slightly cloudy because of the ...
... sterile microfuge tube , dissolve the crude oligonucleotide ( usually 1-10 nmoles ) in 1 ml of sterile , filtered H2O ( Milli - Q or equivalent ) . Vortex the solution thoroughly . The solution is often slightly cloudy because of the ...
Page 11-29
... sterile 50 - ml polypro- pylene tube . Repeat step 1c . 2 3. Add 10 ml of sterile , filtered H2O ( Milli - Q or equivalent ) to the barrel and push it slowly through the column with the plunger . Repeat step 1c . Repeat the water wash ...
... sterile 50 - ml polypro- pylene tube . Repeat step 1c . 2 3. Add 10 ml of sterile , filtered H2O ( Milli - Q or equivalent ) to the barrel and push it slowly through the column with the plunger . Repeat step 1c . Repeat the water wash ...
Page 11-37
... sterile pasteur pipette . Equilibrate the slurry of Bio - Gel P - 60 supplied by the manufac- turer in 10 volumes of 10 mM Tris Cl ( pH 8.0 ) , 0.1 % SDS . Place a sterile glass - wool plug in a sterile pasteur pipette as shown in ...
... sterile pasteur pipette . Equilibrate the slurry of Bio - Gel P - 60 supplied by the manufac- turer in 10 volumes of 10 mM Tris Cl ( pH 8.0 ) , 0.1 % SDS . Place a sterile glass - wool plug in a sterile pasteur pipette as shown in ...
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