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:10/18/2014)... The report "1,6-Hexanediol Market by Application ... 2019", analyzes the global market on the basis ... industry details such as the key drivers, current ... , Browse 81 Market Data Tables and 29 ... on “1,6-Hexanediol Market by Application and by Geography ...
(Date:10/18/2014)... The Asian Electronic Medical Record (EMR) market report defines and ... of revenue. The electronic medical record market in the Asian ... at a CAGR of 9.4% from 2013 to 2018. , ... (EMR) market, to get an idea of the in-depth analysis ... the market in the same region, and is supported by ...
(Date:10/18/2014)... (PRWEB) October 18, 2014 The ... Stress, Holter Monitor), by End-user (Hospitals, Home/Ambulatory, Research ... (Single, 3-6, 12) -Global Forecast to 2020” provides ... challenges, opportunities, current market trends, and strategies impacting ... estimates and forecasts of the revenue and share ...
(Date:10/17/2014)... 17, 2014 For the seventh ... as published in today’s print issue of Science ... Top Employers Survey polled employees in the biotechnology, ... best employers. Rankings were determined based on ... rate their companies on 23 characteristics, including financial ...
Breaking Biology Technology:1,6-Hexanediol Market Projected to Reach $916.1 Million at a CAGR of 8.3% by 2019 – Report by MarketsandMarkets 21,6-Hexanediol Market Projected to Reach $916.1 Million at a CAGR of 8.3% by 2019 – Report by MarketsandMarkets 31,6-Hexanediol Market Projected to Reach $916.1 Million at a CAGR of 8.3% by 2019 – Report by MarketsandMarkets 41,6-Hexanediol Market Projected to Reach $916.1 Million at a CAGR of 8.3% by 2019 – Report by MarketsandMarkets 5The Asian Electronic Medical Record (EMR) market is estimated to grow to $2,328.0 million by 2018 - New Report by MicroMarket Monitor 2The Asian Electronic Medical Record (EMR) market is estimated to grow to $2,328.0 million by 2018 - New Report by MicroMarket Monitor 3The Asian Electronic Medical Record (EMR) market is estimated to grow to $2,328.0 million by 2018 - New Report by MicroMarket Monitor 4The Asian Electronic Medical Record (EMR) market is estimated to grow to $2,328.0 million by 2018 - New Report by MicroMarket Monitor 5The Electrocardiograph (ECG) Market Is Expected to Reach $5,435 Million By 2020 - New Research Report by MarketsandMarkets 2The Electrocardiograph (ECG) Market Is Expected to Reach $5,435 Million By 2020 - New Research Report by MarketsandMarkets 3The Electrocardiograph (ECG) Market Is Expected to Reach $5,435 Million By 2020 - New Research Report by MarketsandMarkets 4The Electrocardiograph (ECG) Market Is Expected to Reach $5,435 Million By 2020 - New Research Report by MarketsandMarkets 5DuPont Named Top Employer by Science Magazine 2DuPont Named Top Employer by Science Magazine 3
... Professor Dr. Manfred T. Reetz honored ... Elsevier , a world-leading provider of ... announces that leading German scientist, Professor Dr. Manfred ... and Hans Meerwein Research Professor at Philipps-Universität, Marburg, ...
... 2012 PRO-DEX, INC. (Nasdaq: PDEX ) ... business, operating under the name Pro-Dex Astromec and located ... of the asset purchase agreement, executed on February 27, ... Astromec, consisting of inventory, equipment and intangibles, and excluding ...
... 2012 Medstreaming, a leading provider of medical IT ... for its All in One EMR/PACS/Practice Management platform at ... Analytic-Based EMR/PACS platform will enable Medstreaming EMR customers to ... data in any format, including DICOM. ...
Cached Biology Technology:Leading German Researcher Awarded Elsevier's 2011 Tetrahedron Prize 2Leading German Researcher Awarded Elsevier's 2011 Tetrahedron Prize 3Pro-Dex, Inc. Announces the Sale Of Its Motor Business 2Medstreaming Launches Web-Based Analytics for All in One EMR/PACS/Practice Management Platform at HIMSS 2Medstreaming Launches Web-Based Analytics for All in One EMR/PACS/Practice Management Platform at HIMSS 3
(Date:10/15/2014)... SHELTON, Conn. , Oct. 15, 2014 NXT-ID, Inc. ... a biometric authentication company focused on the growing mobile commerce market ... NASDAQ MarketSite in Times Square on Monday October 13 th . ... with CTO David Tunnel and angel investor Mr. ... Gino Pereira , CEO of NXT-ID thanked his investors and employees ...
(Date:10/14/2014)... millions of years since T. rex took ... University scientists is breathing life back into dinosaurs using ... The research has important implications for how dinosaurs used ... the sense of smell and cool their brains. ... doctoral student Jason Bourke, lead author of the new ...
(Date:10/14/2014)... CA – October 14, 2014 – Scientists from The ... from the National Institutes of Health (NIH) to lead ... virus-induced hemorrhagic fever disease in Africa. The study aims ... why some patients die, while others survive the inflection. ... understand the basic mechanism of how Lassa fever virus ...
Breaking Biology News(10 mins):Photo Release: NXT-ID Inc. Rings Opening Bell of the NASDAQ Stock Market October 13th 2Photo Release: NXT-ID Inc. Rings Opening Bell of the NASDAQ Stock Market October 13th 3Dinosaur breathing study shows that noses enhanced smelling and cooled brain 2Dinosaur breathing study shows that noses enhanced smelling and cooled brain 3Scripps Research Institute team receives $6.6 million to investigate deadly Lassa virus 2
... associated with an increased risk of hypertension in women, ... sugared or diet colas, according to a study in ... million people in the United States have hypertension, and ... the article. Hypertension is a major risk factor for ...
... by the effects of an intense drought, otherwise harmless ... marsh in the Southeast in recent years. , Periwinkle ... happily with salt marsh. But the drought, which lasted ... such as cordgrass, or Spartina alterniflora, so extensively that ...
... team of scientists in France and the United States ... that enable Rickettsia conorii, the Mediterranean spotted fever pathogen, ... the cells of its host could lead to new ... Rocky Mountain spotted fever and typhus. , Pascale ...
Cached Biology News:No link found between caffeine intake and development of hypertension in women 2Research: Snails were overlooked contributors to marsh destruction 2Research: Snails were overlooked contributors to marsh destruction 3How Rickettsial pathogens break into cells 2
Certified U.S. origin. Bovine Serum:, *Certified U.S. origin. Available raw, nonsterile filtered, or sterile filtered. , , *Sterility tested , , *Low Hemoglobin levels , , *Mycoplasma testing upon...
...
12(S)-HETE-d8 is used as an internal standard for the quantification of 12(S)-HETE by stable isotope dilution mass spectrometry....
Full-length cDNA 5-PRIME end of clone CS0DE013YM09 of Placenta of Homo sapiens (human). [Source:Uniprot/SPTREMBL;Acc:Q86TS7] Antigen: Recombinant Protein Epitope Signature Tag (PrEST)....
Biology Products: