Our blogs and videos choice: plasmid dna purification

JoVE: Electroeluting DNA Fragments (Video Protocol)

Purified DNA fragments are used for different purposes in Molecular Biology and they can be prepared by several procedures. Most of them require a previous electrophoresis of the DNA fragments in order to separate the band of interest. Then, this band is excised out from an agarose or acrylamide gel and purified by using either: binding and elution from glass or silica particles, DEAE-cellulose membranes, "crush and soak method", electroelution or very often expensive commercial purification kits. Thus, selecting a method will depend mostly of what is available in the laboratory. The [more...]

Date: 2010-09-05 22:00:00

Blog posts (24) | Videos (26)
 


Bio Saga Blog: 5-Day Hands-on Workshop on Molecular Biotechnology ...

Recombinant DNA Technology: Cutting & pasting of DNA molecules (i.e. restriction digestion, ligation, DNA gel eectrophoresis, and gel extraction.) Bioinfo part: Bioinformatics of DNA database / SequenceAnalysis: Pairwise sequence alignment, Multiple sequence alignment, Pattern search.Tools: BLAST, CLUSTALW/X, CLC Bio Main Workbench Genetic Engineering: Transformation and plasmid purification, cloning and sub-cloning, amplification of DNA fragments by polymerase chain reaction (PCR). Bioinfo part: Designing of PCR primers In-silico cloning Searching for homologous and paralogous [more...]

Date: 2010-09-29 05:41:00


Endofree Plasmid DNA purification using EMD Fractogel DMAE -650(M)

Precipitate DNA with 0.6-1 vol isopropanol. 10, Carefully pour off as much supernatant as possible, then spin down DNA pellet. 11. Wash pellet with 70-80% ethanol, dissolve DNA in TE or water. 12. Regenerate column by successively applying 1 M NaOH, 2-3 M NaCl, and then either water or  storage solution (100 mM NaCl, 20% ethanol).  Store column at 4 ̊C. P1: 6.1g Tris base 3.7g EDTA•2H2O add water to ~800 ml adjust pH to 8.0 with HCl. Add 100mg of RNase A that had been dissolved in 10ml TE and boiled 15min to inactivate contaminating DNase. Add water to 1 L. P2: 8g NaOH 10g SDS [more...]

Date: 2010-09-25 01:57:59


Plasmid DNA Recovery Using Size-Exclusion and Perfusion ...

Finally, a second size-exclusion chromatography step was carried out to purify the plasmid DNA from other small molecular-weight contaminants. Analytical methods proved that the purified plasmid DNA had a purity of 95% after Sephacryl S1000. Plasmid identity was confirmed by restriction enzyme digestion. Biological activity of the purified plasmid was confirmed in vivo; immunized mice developed a positive antibody response against all HCV structural antigens. This procedure offers an alternative to traditional methods that use organic reagents, mutagenic and toxic compounds, and [more...]

Date: 2008-09-01 04:00:00


Plasmid DNA and transformation questions?

The plasmid purification procedure relies on three main steps to produce a bacterial lysate containing small nucleic acids. The first step uses glucose-Tris-EDT A (GTE). The second uses NaOH/SDS and the third uses sodiumb acetate. [more...]

Date: 2010-02-20 00:17:23


Role of Prostanoid DP Receptor Variants in Susceptibility to ...

Plasmid DNA was purified with the Qiagen plasmid purification system. The effects of constructs (1.5 μg) bearing PTGDR haplotypic variants were studied in transiently transfected A549 cells, grown to 80 percent confluence in six-well [more...]

Date: 2004-10-21 05:00:00


Plasmid DNA extraction from e.coli for digestion mapping clony ...

Research news, bioproducts, protocols, reagents, forums Plasmid DNA extraction from e.coli for digestion mapping clony screening Plasmid DNA extraction from e.coli for digestion mapping clony screening: This protocol utilize the Qiagen buffer P1, P2 and P3 from Qiagen plasmid purification kit. RNase must be added before use. It is cost efficient since it doesn't use expensive plasmid purification columns. This protocol is only intended for screening positive clony. Once positive clonies are determined, purification from bacterial overnight culture can be done using plasmid [more...]

Date: 2006-08-30 07:00:00


The Purification of Plasmid DNA for Clinical Trials Using Membrane ...

ABSTRACTMembrane chromatography offers a good solution to the challenge of developing an efficient chromatographic procedure for plasmid DNA purification. The large convective pores of anion exchange membranes allow plasmid DNA to access all the anionic binding sites of the membrane at high flow rates. Here we demonstrate that the pIDKE2 plasmid can be purified from a recombinant Escherichia coli lysate using a Sartobind D membrane combined with size exclusion chromatography to render material with 95% purity and an average yield of 50%. This process yields therapeutically suitable plasmid [more...]

Date: 2010-02-01 05:00:00


Plasmid double locus sequence typing for IncHI2 plasmids, a ...

Plasmid double locus sequence typing for IncHI2 plasmids, a subtyping scheme for the characterization of IncHI2 plasmids carrying extended-spectrum β-lactamase and quinolone resistance genes Aurora García-Fernández and Alessandra Carattoli* Department of Infectious, Parasitic and Immune-Mediated Diseases, Istituto Superiore di Sanità, Rome, Italy *Corresponding author. Tel: +39-06-49903128; Fax: +39-06-49387112; E-mail: alecara{at}iss.it Received October 9, 2009. Revision requested January 25, 2010. Revision received February 26, 2010. Accepted March 4, 2010. Abstract Objectives IncHI2 [more...]

Date: 2010-03-31 07:00:00


Protostadienol synthase from Aspergillus fumigatus: Functional ...

AGCTATCCAAATTAC-30) with 1 lL of plasmid DNA containing the full length cDNA encoding A. fumigatus OSPC as the template. Phusion DNA polymerase (Finnzymes) was used with dNTP. (0.2 mM) in a final volume of 50 lL, according to the manufac- ... MonoFas DNA purification kit (GL Sciences). The second PCR was performed with 10 lL of this fragment as an anti-sense primer, and 1 lL (20 pmol) of the N-terminal primer (AfuOSPC-N-Spe, 50-. CTTGCCTACTAGTATGGCGACAGACAGCAGCATG- 30) [more...]

Date: 2010-01-06 08:00:00


Miniprep plasmid DNA purification using diatomaceous earth or ...

9. Wash metrix twice with 0.35 ml of PE (0 mM Tris; 100 mM NaCl; 2.5 mM EDTA; 55% (v/v) Ethanol; pH 7.5). 10. Allow the packed matrix to completely dry by spinning for another 1~5 minutes. 11, Add 50 ul EB or water, spin to elute [more...]

Date: 2010-09-05 20:19:25


New ligands for plasmid DNA affinity purification

Molecular therapies and vaccines based on purified plasmid DNA (pDNA) are increasingly being established as promising alternatives to medical classical treatments. The expected wide application of these therapies requires the large-scale production and purification of plasmids. Recent years, had witness the development of new methods for plasmid purification, based on aqueous two-phase systems (ATPS) offering additional advantages like technical simplicity and the use of inexpensive equipment and chemicals. This project is going to study the molecular recognition mechanism and binding [more...]

Date: 2010-06-05 13:04:07


solutions manual to introduction to particle technology ...

High-Throughput Solutions for DNA Purification Using MagneSil … INTRODUCTION. MagneSilâ„¢ technology was developed specifically to meet the changing … small particle size allows rapid movement in a magnetic field, resulting … plasmid DNA in <2 hours with no manual interventions. Unique magnetic … http://www.promega.com/pnotes/75/8554_07/8554_07.pdf * pdf N07848_ver2.5 MagMAX Express User Manual Principle of the MagMAXâ„¢ Express magnetic particle processor . ….. Chapter 3: Introduction to the MagMAXâ„¢ Express …. The operating principle employed is MPP (inverse [more...]

Date: 2010-10-13 07:08:10


(IUCr) Mammalian cell expression, purification, crystallization ...

was amplified by PCR from plasmid DNA (using 5'-TGGTACCGCCACCATGGCAAGACAAGGCTGTCTC-3' and 5'-ACCGGTAATCTCGCTCTCATATGT-3' primers) and the reaction product was cut with Asp718I and AgeI restriction enzymes and cloned into the plasmid [more...]

Date: 2010-08-31 07:00:00


Lab Tech / Research Associate (Torrey Mesa) | PCR Jobs

Candidate must be familiar with basic molecular biology techniques such as DNA purification, DNA gel electrophoresis, restriction digestion, PCR including primer design, plasmid construction, DNA sequence analysis software and bacterial transformation. Candidate will also assist with sample receipt and tracking, maintain inventory and equipment, and perform general lab maintenance and cleaning to include handling of biological waste. Education/ Skills/Attributes Required: · Must be a US Citizen. · BS degree in biology/biochemistry with 0-3 years industrial experience. · Experience with [more...]

Date: 2010-10-12 19:15:31


Rapid, nondenaturing RNA purification using weak anion-exchange ...

We present a simple and fast method for large-scale purification of RNA oligonucleotides suitable for biochemical and structural studies. RNAs are transcribed in vitro with T7 RNA polymerase using linearized plasmid DNA templates. [more...]

Date: 2010-01-25 15:06:07


Most Important Discoveries in Vaccinology Part II Good Tips ...

You are here: Home / Vector / Most Important Discoveries in Vaccinology Part II Good Tips Most Important Discoveries in Vaccinology Part II Polysaccharides, Bacterial polysaccharide The way this will open the pre World War II, but the operation the immunological effect of the combination of bacterial polysaccharide with proteins has happened only recently. The 80′s saw the implementation of this technology coma Hib vaccine as a first conjugate being licensed for infants in 1990; A reminder is not necessary for the spectacular success of the Hib vaccine, which promises eradicate the disease [more...]

Date: 2010-10-10 19:00:01


HIV Vaccine | Biology in Action

“The prime is a plasmid DNA vaccine, PENNVAX™-G, and the boost is a virus vector vaccine, Modified Vaccinia Ankara-Chiang Mai Double Recombinant (MVA-CMDR). Together, the vaccines are designed to deliver a diverse mixture of antigens for HIV-1 subtypes A, B, C, D and E”. Taken separately, the strategies have been proven safe. Several precautions have to be taken because the study is the use of different HIV antigens during the priming (A-D) and boosting A/E. The DNA vaccine was designed at the University of Pennsylvania and licensed by Inovio Pharmaceuticals. The vaccine is based off a [more...]

Date: 2010-10-12 13:07:22


(IUCr) Cloning, purification, crystallization and preliminary X ...

Cloning, purification, crystallization and preliminary X-ray diffraction of the OleC protein from Stenotrophomonas maltophilia involved in head-to-head hydrocarbon biosynthesis. Janice A. Frias,a Brandon R. Goblirsch,a Lawrence P. Wacketta ... The DNA was cloned into a pET30b vector (Novagen, Madison, Wisconsin, USA) containing a C-terminal His tag. The recombinant plasmid was transformed into Escherichia coli BL21 (DE3) pLysE One Shot cells (Invitrogen) for expression. [more...]

Date: 2010-08-28 07:00:00


Plasmid DNA Extraction (Miniprep) Video | DnaTube.com - Scientific ...

NOTICE: YOU NEED TO FILL YOUR PROFILE PROPERLY TO BE ABLE TO DOWNLOAD VIDEO. LOOK BELOW TO SEE THE NECESSARY INFORMATION TO BE FILLED. Your acedemic background information is missing. Please click here to fill this information. Your occupation information is missing. Please click here to fill this information. Your current school and company information is missing. Please click here to fill this information. Your current "about me" information is missing. Please click here to fill this [more...]

Date: 2010-04-27 07:00:00


Plasmid DNA Extraction (Midiprep) Video | DnaTube.com - Scientific ...

NOTICE: YOU NEED TO FILL YOUR PROFILE PROPERLY TO BE ABLE TO DOWNLOAD VIDEO. LOOK BELOW TO SEE THE NECESSARY INFORMATION TO BE FILLED. Your acedemic background information is missing. Please click here to fill this information. Your occupation information is missing. Please click here to fill this information. Your current school and company information is missing. Please click here to fill this information. Your current "about me" information is missing. Please click here to fill this [more...]

Date: 2010-05-18 07:00:00


Plasmid v Genomic DNA Extraction:The Difference | Bitesize Bio

If you want to isolate plasmid DNA, you crack your cells open and carry out a miniprep, trying very hard not to get any contaminating genomic DNA in [more...]

Date: 2009-04-16 08:49:09


Plasmid Isolation - Haywood, Community, College, Biotechnology ...

4 Genetic Engineering con... Frontiers of Biomedical Engineering (BENG 100) Professor Saltzman continues his ... 4 Genetic Engineering con... Frontiers of Biomedical Engineering (BENG 100)Professor Saltzman continues his [more...]

Date: 2008-07-10 07:00:00


Vical signs deal to make HIV vaccines - CNBC

Some companies have figured out that if you get a little creative, it can go a long [more...]

Date: 2010-10-07 11:12:00



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