Molecular Cloning: A Laboratory Manual, Book 2 |
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Page 13-24
... step 2. If it is not possible to avoid restriction enzymes that generate blunt ends , the ligation ( step 2 ) should be carried out in the presence of substances that increase macromolecular crowding ( e.g. , polyethylene glycol ...
... step 2. If it is not possible to avoid restriction enzymes that generate blunt ends , the ligation ( step 2 ) should be carried out in the presence of substances that increase macromolecular crowding ( e.g. , polyethylene glycol ...
Page 14-31
... step 6 . It is important to use water rather than TE as the diluent so as to avoid altering the concentration of Mg ** in the polymerase chain reaction . 3. Set up a series of amplification reactions as described on page 14.18 ( DNA ...
... step 6 . It is important to use water rather than TE as the diluent so as to avoid altering the concentration of Mg ** in the polymerase chain reaction . 3. Set up a series of amplification reactions as described on page 14.18 ( DNA ...
Page 15-25
... step 15. Add 1 μl of gel - loading buffer ( see Appendix B ) to each aliquot , and load the contents of each tube into the wells of an agarose gel containing ethidium bromide ( 0.5 μg / ml ) . The wells at the sides of the gel should ...
... step 15. Add 1 μl of gel - loading buffer ( see Appendix B ) to each aliquot , and load the contents of each tube into the wells of an agarose gel containing ethidium bromide ( 0.5 μg / ml ) . The wells at the sides of the gel should ...
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Common terms and phrases
agarose gel aliquots amplification annealing antibody bacterial bacteriophage M13 bacteriophage T4 bacteriophage T4 DNA buffer carried cDNA clones cDNA libraries cells centrifugation at 12,000g Chapter cleavage cleaved cleotide codon coli DNA polymerase concentration containing deletions denatured digestion dithiothreitol DNA ligase DNA sequencing DNAase dNTPs double-stranded DNA EDTA EDTA pH 8.0 efficiency ethanol exonuclease formamide fusion proteins gel electrophoresis gene genomic DNA H₂O hybridization Incubate inserted Klenow fragment ligation linear linkers method microfuge tube minutes at 4°C molecules mRNA mutagenesis mutagenic oligonucleotide mutations Natl nitrocellulose nitrocellulose filters Nucleic Acids nucleotides oligonucleotide phagemid plaques plasmid plasmid DNA plate polymerase chain reaction prepared primer probes purified radioactivity radiolabeled reaction mixture recombinant remove restriction enzyme reverse transcriptase room temperature samples screening sequencing reactions single-stranded DNA solution step Store strand of cDNA synthesis target DNA target sequence template DNA termini transfer Tris Cl pH vector