Lysis of colonies and DNA binding assay to filter membranes
This protocol describes how to release DNA from a clone carrying a recombinant plasmid and immobilize it on a nitrocellulose filter or nylon membrane, a method
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Preparation of λ phage proto-species experiments with small quantities of liquid cultures
High-titer λ phage progenitors can be prepared from small quantities of liquid cultures. This method was first used by Leder et al. (1977). In general, i
Preparation of λ phage DNA by double restriction enzyme cleavage as a cloning vector
Replacement vectors (e.g., λ2001, λDASH, EMBL series, Charon 34, Charon 35, and Charon 40) contain a series of restriction sites that are aligned
Preparation of single-stranded DNA using phage vectors
Phage plasmids skillfully combine the characteristics of plasmids and filamentous phages. In addition to the basic features, these plasmids are usually high cop
Screening of DNA binding proteins using λ phage libraries
Synthetic double-stranded oligonucleotides can be used to screen cDNA expression libraries constructed in λ phage to identify clones corresponding to spe
λ Purification of phage arms (by sucrose density gradient centrifugation) experiments
Unlike insertion vectors, replacement vectors contain a filler fragment in the middle of the genome, which must be removed in order to accommodate the foreign D
Construction of genomic DNA libraries using mucoid vectors
The steps for constructing genomic DNA libraries in λ phage are essentially the same as in mucosal vectors. In both systems, eukaryotic DNA is ligated to vecto
Partial cleavage (pre-reaction) of eukaryotic DNA for genomic libraries
Regardless of the base composition and sequence of high-molecular-quality DNA, it can be fragmented in a semi-random fashion by hydrodynamic shearing. However,
Recombinant M13 phage cloning analysis
In this protocol, when the exogenous DNA is larger than 200~300 nucleotides, it can be identified by gel electrophoresis analysis of the single-stranded DNA rel