Plasmid DNA preparation by the toothpick method in small quantities
Colonies picked with a toothpick from agar medium can be used directly to prepare plasmid DNA. the resulting closed-loop DNA often has too many impurities to be
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Plasmid DNA purification by polyethylene glycol precipitation assay
This method was first devised by Richaed Treisman (ICRF, London, UK) with reference to the work of Lis, who was the first to use polyethylene glycol (PEG) to is
Chemiluminescent Dideoxy DNA Sequencing Assay
Standard double deoxy DNA sequencing can be easily and efficiently detected using non-isobaric labeled chemiluminescence. In the sequencing reaction, biotin-lab
Establishment of randomized overlapping DNA insertion libraries
The following protocol describes the starting steps of birdshot sequencing, from shearing of target DNA to construction of a DNA fragment library with M13 phage
Enzymatic digestion of DNA fragments
Restriction endonucleases are a class of DNA hydrolases that recognize specific nucleotide sequences in double-stranded DNA molecules, and the discovery and app
Ligation of plasmid and target DNA in low melting point agarose experiment
The most time-consuming step of the plasmid cloning operation is the purification of the desired sized fragments of exogenous and plasmid DNA by electrophoresis
Agarose gel electrophoresis of DNA
DNA electrophoresis can be used to (1) separate DNA fragments of different sizes, (2) identify target DNA fragments, and (3) purify and recover DNA fragments.
Dephosphorylation of plasmid DNA
Removal of the phosphate group at the 5' end prevents self-ligation and cyclization of plasmid DNA. In an in vitro ligation reaction, DNA ligase forms a phospho
Total DNA quality and digestion assay
The objectives of the experiment are to understand and master the methods of detecting the quality of DNA as well as the methods of DNA quantification; to under