Alkaline lysis

Alkaline lysis

Alkaline lysis is a method used in molecular biology to break cells open to isolate plasmid DNA or other cell components such as proteins. Bacteria containing the plasmid of interest is first grown, then lysed with a strong alkaline buffer consisting of a detergent sodium dodecyl sulfate (SDS) and a strong base sodium hydroxide. The detergent breaks the membrane's phospholipid bilayer and the alkali denatures proteins involved in maintaining the structure of the cell membrane. Through a series of steps involving agitation, precipitation, centrifugation, and the removal of supernatant, cellular debris is removed and the plasmid is isolated and purified. The protocol may vary slightly from lab to lab.

Also, alkaline lysis is sometimes used to extract plant genetic material. The plant cells are subjected to a strongly alkaline solution containing a detergent (usually a zwitterionic or nonionic detergent such as Tween 20), and the mixture is incubated at high temperature. This method is not used as often due to the sodium hydroxide's tendency to damage genetic material, reducing DNA fragment size.

References

* [http://lifesciences.asu.edu/resources/mamajis/alkaline/alkaline.html Alkaline Lysis at Mama Ji's Molecular Kitchen]
* [http://www.pubmedcentral.nih.gov/articlerender.fcgi?tool=pubmed&pubmedid=388356 A rapid alkaline extraction procedure for screening recombinant plasmid DNA] by H. C. Birnboim and J. Doly


Wikimedia Foundation. 2010.

Игры ⚽ Нужно сделать НИР?

Look at other dictionaries:

  • Plasmid preparation — Plasmid miniprep. 0.8% agarose gel ethidium bromide stained. A plasmid preparation is a method used to extract and purify plasmid DNA. Many methods have been developed to purify plasmid DNA from bacteria. These methods invariably involve three… …   Wikipedia

  • Mycobacterium ulcerans — Scientific classification Kingdom: Bacteria Phylum: Actinobacteria Order: Actinomycetales …   Wikipedia

  • Triton X-100 — Other names polyethylene glycol p (1,1,3,3 tetramethylbutyl) phenyl ether, octyl phenol ethoxylate, polyoxyethylene octyl phenyl ether, 4 oct …   Wikipedia

  • List of molecular biology topics — This is a list of topics in molecular biology. See also list of biochemistry topics. NOTOC # 3 end 3 flanking region 5 end 5 flanking region 5 ribose 3 A acrylamide gels adenine adenosine deaminase deficiency adenovirus agarose gel… …   Wikipedia

  • Plasmid — Figure 1: Illustration of a bacterium with plasmid enclosed showing chromosomal DNA and plasmids. In microbiology and genetics, a plasmid is a DNA molecule that is separate from, and can replicate independently of, the chromosomal DNA …   Wikipedia

  • Comet assay — The Single Cell Gel Electrophoresis assay (also known as comet assay) is an uncomplicated and sensitive technique for the detection of DNA damage at the level of the individual eukaryotic cell. It was first described by Singh et al. in 1988. It… …   Wikipedia

  • test — 1. To prove; to try a substance; to determine the chemical nature of a substance by means of reagents. 2. A method of examination, as to determine the presence or absence of a definite disease or of some substance in any of the fluids, tissues,… …   Medical dictionary

  • poison — poisoner, n. poisonless, adj. poisonlessness, n. /poy zeuhn/, n. 1. a substance with an inherent property that tends to destroy life or impair health. 2. something harmful or pernicious, as to happiness or well being: the poison of slander. 3.… …   Universalium

  • reaction — 1. The response of a muscle or other living tissue or organism to a stimulus. 2. The color change effected in litmus and certain other organic pigments by contact with substances such as acids or alkalies; also the property that such substances… …   Medical dictionary

  • protein — proteinaceous /proh tee nay sheuhs, tee i nay /, proteinic, proteinous, adj. /proh teen, tee in/, n. 1. Biochem. any of numerous, highly varied organic molecules constituting a large portion of the mass of every life form and necessary in the… …   Universalium

Share the article and excerpts

Direct link
… Do a right-click on the link above
and select “Copy Link”