Ligation experiments of λ phage arms to exogenous genomic DNA fragments
When a λ phage arm is attached to an exogenous genomic DNA fragment, two parameters must be taken into account: the molar ratio of the phage arm to the potenti
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Phage DNA hybridization experiments on filter membranes
Filter membranes carrying immobilized DNA derived from phage spots can be screened by in situ hybridization with 32P-labeled probes. The technique is robust, hi
Preparation of small quantities of yeast DNA
Yeast DNA can be prepared by digesting the cell wall and lysing the resulting protoplasts with SDS, and several milligrams of yeast DNA can be reproducibly prep
Detection of DNA in agarose gels
Nucleic acids in agarose gels can be detected by staining under UV light at a wavelength of 300 nm. Two methods for staining nucleic acids in agarose gels are i
Phage DNA transfer experiments from phage spots to filter membranes
A method for identifying and isolating specific recombinants from λ phage libraries was established early in the history of molecular cloning by Bemon and Davi
Rapid analysis of λ phage isolates (purification of λ DNA from liquid cultures) Experiments
As described in this scheme, λ phage DNA can be easily purified from liquid cultures. In contrast, the previous scheme describes a method for purifying λ phag
Oligonucleotide 5' terminal phosphorylation assay
The following describes a reaction for labeling a 10 pmol high specific activity oligonucleotide. Labeling of different amounts of oligonucleotides can be accom
Isolation of genomic DNA fragment ends in high-capacity vectors (small vector PCR)
The genes of many eukaryotes contain far more DNA than a single recombinant can hold. This is especially true for most chromosomes. Therefore, it is necessary t
Southern blotting (capillary transfer of DNA to a membrane)
Genomic DNA samples are prepared by digestion with one or more restriction endonucleases, and the digested fragments are separated by electrophoresis on a stand