Navigation Links
Basic Plasmid DNA Isolation Protocol

PLG can be used in small and large scale protocols based on alkaline lysis procedure.(1,2,3)

The basic protocol given here works with E. coli cultures grown for 1214 hours in up to 500 ml of amplified or non-amplified LB or DYT, or in 250 ml Terrific Broth.

  1. Pellet bacteria from the culture at 10,000 x g for 5 minutes at room temperature.
  2. Resuspend bacterial pellet in a total of 15 ml 25 mM Tris-Cl/10 mM EDTA (pH 8.0). Pipet up and down or vortex as necessary to fully resuspend the bacteria.
  3. Add 15 ml room temperature 0.2 N NaOH/1.0% SDS to the suspension. Mix thoroughly by repeated gentle inversion. Do not vortex.
  4. Add 15 ml ice-cold 7.5 M Ammonium Acetate to the lysate. Mix thoroughly by repeated gentle inversion. Do not vortex.
  5. Centrifuge at 15,500 x g for 30 minutes at 4C.
  6. Recover resulting supernatant and divide it between 2 pre-spun (1500 x g for 23 minutes) PLG 50 ml Heavy tubes.
  7. Add 20 ml Phenol-Chloroform-Isoamyl Alcohol (PCI, 25:24:1) to the sample in each PLG 50 ml Heavy tube and mix thoroughly by repeated inversion. Do not vortex. Centrifuge at 1500 x g for 5 minutes to separate the phases.
  8. Recover resultant aqueous phases to fresh pre-spun PLG 50 ml Heavy tubes and repeat step 7 above.
  9. Recover resultant aqueous phases to fresh pre-spun PLG 50 ml Heavy tubes. Add 20 ml Chloroform-Isoamyl Alcohol (CI, 24:1) and extract as in step 7 above.
  10. Recover resultant aqueous phases to suitable tubes and
    '"/>


Source:


Page: All 1 2

Related biology technology :

1. Basic Plasmid DNA Isolation Protocol
2. Array Analysis: The Basics
3. Northern Analysis: The Basics
4. Nuclease Protection Assays: The Basics
5. Efficiently Insert Unique Restriction Sites into Plasmid Vectors
6. High-Efficiency Site-Directed Mutagenesis of Large Plasmid Vectors
7. Simple Purification of DNA from Plasmid Minipreps, PCR Amplifications and Agarose Gels
8. Generate High-Quality, Directional Plasmid cDNA Libraries
9. New Reporter Plasmids for Studying Interferon-Stimulated Signal Transduction Pathways
10. New Fusion Trans-Activator Plasmids for Studying Signal Transduction Pathways
11. Comparison of Growth Techniques and Media for the Purpose of Plasmid Isolation from E. coli Using the Eppendorf Perfectprep Plasmid Mini Kit
Post Your Comments:
*Name:
*Comment:
*Email:
TAG: Basic Plasmid DNA Isolation Protocol

(Date:9/30/2014)... , Sept. 30, 2014 FemCap Inc., ... health announces its receipt of a patent No: US, ... cup has also received FDA clearance in the U.S., ... the CE in Europe. This patent is related to its ... this issuance, the company is releasing FemmyCycle Teen, a ...
(Date:9/30/2014)... SoundConnect, an award ... dedicated to supporting business communication solutions, including business ... a carrier services specialist has joined ... connected and productive at all times. This growth ... businesses to cloud communications services. On a ...
(Date:9/30/2014)... 2014 Back in the 1970’s, many companies ... per employee, which included space for circulation and file storage. ... Howell, partner at Nidea Corporate Real Estate / ITRA ... the year 2000, however, 250 square feet per employee ... come down to 175 to 225 square feet, and over ...
(Date:9/29/2014)... Sept. 29, 2014  Ten of the most ... cartilage repair, regeneration, allograft reconstruction and research and ... world gathered to discuss the future of cartilage ... Regenerative Medicine symposium, Cartilage Regeneration: State of ... Orthopedic Biosurgery, Minced Juvenile Allograft, and State of ...
Breaking Biology Technology:The FemmyCycle 2SoundConnect Expands Carrier Services Division 2ITRA Global Reports on How Companies Are Squeezing More Employees into Less Office Space 2ITRA Global Reports on How Companies Are Squeezing More Employees into Less Office Space 3World Renowned Experts Discuss Future Of Cartilage Regeneration 2World Renowned Experts Discuss Future Of Cartilage Regeneration 3
... Ill., June 30 Hospira, Inc. (NYSE: ... company, today announced agreements with MedAssets on intravenous (I.V.) ... its agreement for infusion devices. The five-year MedAssets agreements ... solutions and equipment, and July 1, 2009, for infusion ...
... needs, clinical uses and funding sources , , ROCKVILLE, ... in Rockville, Md., announced today that former Secretary of the ... board of directors. , , "I am excited ... battlefield and in emergency rooms throughout the US. I am ...
... Limited, the decentralised and point-of-care molecular diagnostics company, announced today ... Veterinary Laboratories Agency to evaluate the performance of the Enigma ... be one of the main threats to cattle (1), affecting ... closely related to classical swine fever virus, a serious disease ...
Cached Biology Technology:Hospira Announces New Contract With MedAssets for Intravenous Solutions, Renewal of Infusion Devices Agreement 2Former Secretary of Veterans Affairs James Peake to Join the Cellphire Board of Directors 2Enigma Diagnostics Announces Successful Evaluation of the Enigma FL by the UK's Veterinary Laboratories Agency 2Enigma Diagnostics Announces Successful Evaluation of the Enigma FL by the UK's Veterinary Laboratories Agency 3
(Date:9/30/2014)... drinking water may increase the risk of stillbirth ... led by a Boston University School of Public ... journal Environmental Health , compared 1,091 PCE-exposed ... in Cape Cod, Ma., where water was contaminated ... by the installation of vinyl-lined asbestos cement pipes. ...
(Date:9/30/2014)... involving mice and published in the October 2014 issue ... , suggests that just because you can do it, ... of Italian scientists have found that using marijuana in ... system. This damage may result in autoimmune diseases and ... disease and rheumatoid arthritis in adulthood. , "I hope ...
(Date:9/30/2014)... , Sept. 30, 2014  Spectra ... solutions for the biotech and power generation ... new RECONN Bioprocess Manager, an easy-to-use and ... geared towards bioprocess development laboratories. ... disparate bioreactor and analyzer systems. Accessing and ...
Breaking Biology News(10 mins):Contaminated water linked to pregnancy complications, BU study finds 2Adolescent exposure to thc may cause immune systems to go up in smoke 2Spectra Automation Introduces RECONN Bioprocess Manager 2
... blood cell production and function will today be awarded ... Council,s (NHMRC) program grants scheme. The nine ... securing $107 million in program grant funding will be ... the Federal Minister for Mental Health and Ageing, the ...
... Parachin and the secret of her technique is enzymes that ... made from xylose then ethanol production could increase by over ... friendly fuel. Ethanol is manufactured by fermenting sugars from ... the second most common type of sugar found in nature. ...
... -- Software agents on assignment . . . ... organization,s network could be more secure with a system ... Oak Ridge National Laboratory,s Computational Sciences and Engineering Division. ... are being copied, excerpted, changed and stored in various ...
Cached Biology News:$38.4M NHMRC program grants extend cancer and blood cell research 2Enzymes from garden compost could favour bioethanol production 2Story tips from the US Department of Energy's Oak Ridge National Laboratory March 2011 2
... Motic's B3 series of microscopes offer ... microscope required for laboratory work. Equipped with ... Super Contrast (ASC) objectives. The B3 is ... The Siedentopf binocular tube eliminates the need ...
... SYBR Green II RNA gel ... gel stain that has bright fluorescence when ... gels, making it ideal for use with ... scanners or standard UV transilluminators. A 500 ...
... SYBR Green II RNA gel stain ... has bright fluorescence when bound to RNA and ... use with either formaldehyde/agarose or polyacrylamide gels using ... stains 100 minigels. A 500 µL unit size ...
... Glutathion-S-transferases (GST) are a group ... the detoxification processes. The GSTs are ... Pi, Teta and microsomal. The classes ... and s Sample Size: ...
Biology Products: