» 
Arabic Bulgarian Chinese Croatian Czech Danish Dutch English Estonian Finnish French German Greek Hebrew Hindi Hungarian Icelandic Indonesian Italian Japanese Korean Latvian Lithuanian Malagasy Norwegian Persian Polish Portuguese Romanian Russian Serbian Slovak Slovenian Spanish Swedish Thai Turkish Vietnamese
Arabic Bulgarian Chinese Croatian Czech Danish Dutch English Estonian Finnish French German Greek Hebrew Hindi Hungarian Icelandic Indonesian Italian Japanese Korean Latvian Lithuanian Malagasy Norwegian Persian Polish Portuguese Romanian Russian Serbian Slovak Slovenian Spanish Swedish Thai Turkish Vietnamese

definition - Plasmid_preparation

definition of Wikipedia

   Advertizing ▼

Wikipedia

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 steps:

  • Growth of the bacterial culture
  • Harvesting and lysis of the bacteria
  • Purification of plasmid DNA

Contents

  Growth of the bacterial culture

Plasmids are almost always purified from liquid bacteria cultures, usually E. coli, which have been transformed and isolated. Virtually all plasmid vectors in common encode one or more antibiotic resistance genes as a selectable marker (Ex :kanamycin,Ampicillin), which allows bacteria that have been successfully transformed to multiply uninhibited. Bacterials are grown under favourable conditions.

  Harvesting and lysis of the bacteria

When bacteria are lysed under alkaline conditions both DNA and protein are precipitated. Some scientists reduce the concentration of NaOH used to 0.1M in order to reduce the occurrence of ssDNA. After the addition of acetate-containing neutralization buffer the large and less supercoiled chromosomal DNA and proteins precipitate, but the small bacterial DNA plasmids can renature and stay in solution.

  Preparations by size

Kits are available from varying manufacturers to purify plasmid DNA, which are named by size of bacterial culture and corresponding plasmid yield. In increasing order, these are the miniprep, midiprep, maxiprep, megaprep, and gigaprep. The plasmid DNA yield will vary depending on the plasmid copy number, type and size, the bacterial strain, the growth conditions, and the kit.

  Minipreparation

Minipreparation of plasmid DNA is a rapid, small-scale isolation of plasmid DNA from bacteria. It is based on the alkaline lysis method invented by the researchers Birnboim and Doly in 1979. The extracted plasmid DNA resulting from performing a miniprep is itself often called a "miniprep". Minipreps are used in the process of molecular cloning to analyze bacterial clones. A typical plasmid DNA yield of a miniprep is 20 to 30 µg depending on the cell strain.

  Miniprep Protocols

http://www.protocol-online.org/prot/Molecular_Biology/Plasmid/Miniprep/

A miniprep procedure using diatomaceous earth to bind DNA during purification and washing.

  Midipreparation

The starting E. coli culture volume is 15-25 ml of LB broth and the expected DNA yield is 100-350 µg.

  Maxipreparation

The starting E. coli culture volume is 100-200 ml of LB broth and the expected DNA yield is 500-850 µg.

  Megapreparation

The starting E. coli culture volume is 500 ml – 2.5 L of LB broth and the expected DNA yield is 1.5-2.5 mg.

  Gigapreparation

The starting E. coli culture volume is 2.5-5 L of LB broth and the expected DNA yield is 7.5–10 mg.

  Purification of plasmid DNA

Addition of phenol/chloroform can dissolve and denature proteins, like DNase. This is especially important if the plasmids are to be used for enzyme digestion. Otherwise, smearing may occur in enzyme restricted form of plasmid DNA.

  References

Birnboim HC, Doly J (November 1979). "A rapid alkaline extraction procedure for screening recombinant plasmid DNA". Nucleic Acids Res. 7 (6): 1513–23. doi:10.1093/nar/7.6.1513. PMC 342324. PMID 388356. http://nar.oxfordjournals.org/cgi/pmidlookup?view=long&pmid=388356. 


   
               

 

All translations of Plasmid_preparation


sensagent's content

  • definitions
  • synonyms
  • antonyms
  • encyclopedia

Dictionary and translator for handheld

⇨ New : sensagent is now available on your handheld

   Advertising ▼

sensagent's office

Shortkey or widget. Free.

Windows Shortkey: sensagent. Free.

Vista Widget : sensagent. Free.

Webmaster Solution

Alexandria

A windows (pop-into) of information (full-content of Sensagent) triggered by double-clicking any word on your webpage. Give contextual explanation and translation from your sites !

Try here  or   get the code

SensagentBox

With a SensagentBox, visitors to your site can access reliable information on over 5 million pages provided by Sensagent.com. Choose the design that fits your site.

Business solution

Improve your site content

Add new content to your site from Sensagent by XML.

Crawl products or adds

Get XML access to reach the best products.

Index images and define metadata

Get XML access to fix the meaning of your metadata.


Please, email us to describe your idea.

WordGame

The English word games are:
○   Anagrams
○   Wildcard, crossword
○   Lettris
○   Boggle.

Lettris

Lettris is a curious tetris-clone game where all the bricks have the same square shape but different content. Each square carries a letter. To make squares disappear and save space for other squares you have to assemble English words (left, right, up, down) from the falling squares.

boggle

Boggle gives you 3 minutes to find as many words (3 letters or more) as you can in a grid of 16 letters. You can also try the grid of 16 letters. Letters must be adjacent and longer words score better. See if you can get into the grid Hall of Fame !

English dictionary
Main references

Most English definitions are provided by WordNet .
English thesaurus is mainly derived from The Integral Dictionary (TID).
English Encyclopedia is licensed by Wikipedia (GNU).

Copyrights

The wordgames anagrams, crossword, Lettris and Boggle are provided by Memodata.
The web service Alexandria is granted from Memodata for the Ebay search.
The SensagentBox are offered by sensAgent.

Translation

Change the target language to find translations.
Tips: browse the semantic fields (see From ideas to words) in two languages to learn more.

last searches on the dictionary :

4404 online visitors

computed in 0.031s

   Advertising ▼

I would like to report:
section :
a spelling or a grammatical mistake
an offensive content(racist, pornographic, injurious, etc.)
a copyright violation
an error
a missing statement
other
please precise:

Advertize

Partnership

Company informations

My account

login

registration

   Advertising ▼