λ Mass culture (low multiplicity infection) experiments with phages
Mass preparation of λ phage can be obtained by either low-mass infection of bacterial cultures or high-mass infection. Immediately after low magnificatio
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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
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
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
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
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
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
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,
Large-scale preparation of single- and double-stranded M13 phage DNAs
This protocol is mainly used to prepare large quantities of double-stranded DNA from M13 phage, which is often used as a cloning vector in the laboratory, as we