E. Coli Plasmid Vectors: Methods and Applications (Methods in ...
E. Coli Plasmid Vectors: Methods and Applications (Methods in Molecular Biology) E. Coli Plasmid Vectors: Methods and Applications (Methods in Molecular Biology) (archive) E. Coli Plasmid Vectors: Methods and Applications (Methods in Molecular Biology (Clifton, N.J.), V. 235.) Table of Contents The Function and Organization of Plasmids Finbarr Hayes Choosing a Cloning Vector Andrew Preston Escherichia coli Host Strains Nicola Casali Chemical Transformation of E. coli W. Edward Swords Electroporation of E. coli Claire A. Woodall DNA Transfer by Bacterial Conjugation [more...]
Date: 2004-04-04 08:00:00
ERV's latest blog on the FDA paper. One step too far?.
PNAS paper, using standard PCR to ID 'positives' and 'negatives', with insane numbers of bands as a result of non-specific primer binding. Insane numbers. As if this is 1990 and we dont know what 'Real Time PCR' and 'Taqman Probes' are. Im going to ignore that and grant the premise they really have IDed viral sequences. And they arent XMRV. They dont cluster anywhere near XMRV on a phylogenetic tree. Theyre something else entirely. Okay, well, logically it could be contamination from mouse DNA. So they checked for that... by looking for mouse mitochondrial DNA. *blink* I understand [more...]
Date: 2010-08-23 07:00:00
ISPUB - Identification Of Lipase – Producing Psychrophilic Yeast ...
Lambda HindIII was used as DNA marker. After the electrophoresis, gel was stained with ethidium bromide and documented by the Gel Doc 2000 (Bio-Rad). Purification of PCR product was done by using QiAquick Gel Extraction Kit. Then, the purified PCR product obtained was sequenced and submitted for BLAST (Basic Local Alignment Search Tools) at NCBI (National Center for Biotechnological Information). Lipase assay by titrationActivity of lipase was assayed by titration using oil emulsion (polyvinyl alcohol: olive oil; 3:1) as substrate (Arima et al., 1972). The assay medium consists of 5ml oil [more...]
Date: 2010-08-25 00:51:13
DNA sequencing » Blog Archive » DNA Sequencing Gel
Preparation of DNA Sequencing Gel 3. Assemble the gel rig on the sequencer, being careful not to get … 7. Run the gel for 45 minutes. E. Denature the sequencing reactions and prepare them for … DNA sequencing gel Hahn Lab Methods DNA sequencing gel. Acrylamide Urea Gel (100 ml) … if the top of the gel is wet with water and is wrapped tightly in Saran Wrap … How do we Sequence DNA? DNA Sequencing is at the center of the Human Genome Project, which promises to … sequencing gel: That’s exactly what we do to sequence DNA, then … Chapter 8 A. Recombinant DNA Technology PCR has [more...]
Date: 2010-09-28 04:49:52
Thrombosis Journal | Full text | Effect of PlA1/A2 glycoprotein ...
Genomic DNA was prepared from peripheral lymphocytes by the salt precipitation method . The PlA1/A2 alleles of the GP IIIa gene were identified on the basis of MspI enzyme site restriction analysis after amplification of a 476 base pairs GP IIIa fragment (sens amorce 5'-ATA-AGC-TTA-GCT-ATT-GGG-AAG-TGG-TAG-GGC-CTG-3', antisens amorce 5'-CTT-CTG-ACT-CAA-GTC-CTA-ACG-3'). The GP IIIa gene was amplified using the polymerase chain reaction (PCR) method. Each amplification product was verified on an agarose gel. Amplification results in a 476 base pairs (bp) fragment. Digestion was obtained with [more...]
Date: 2008-01-15 08:00:00
Thrombosis Journal | Full text | Expression of sterol regulatory ...
DNA was extracted from frozen cardiac muscle samples. The SCAP 2386A>G genotyping was based on PCR amplification, restriction enzyme analysis and DNA electrophoresis. The DNA samples were amplified by PCR, using the primers 5'-TTGTGCTGCGCGGCCACCTCA-3' and 5'-AGGAGGAAAGGGCAGCCGCAC-3'. PCR was performed in a volume of 50 μl. Cycle conditions were 94°C for 4 min, then 28 cycles of 94°C for 1 min, 64°C for 1 min and 72°C for 1 min, with a final extension step of 5 min at 72°C in a PTC-225 thermal cycler (MJ research, Massachusetts, USA). 10% DMSO was included in PCR reaction. The [more...]
Date: 2009-02-18 08:00:00
Genetic Fingerprinting of Bacillus thuringiensis Isolates by ...
Genetic Fingerprinting of Bacillus thuringiensis Isolates by Randomly Amplified Polymorphic DNA Polymerase Chain Reaction (RAPD-PCR)(Published in Nepal Journal of Science and Technology 10(2009) 97-103)Gyan Sundar Sahukhall, Upendra Thapa Shrestha1, Binod Lekhak2, Anjana Singh2, Viswanath Prasad Agrawal11Research Laboratory for Biotechnology and Biochemistry (RLABB), Kathmandu, Nepal. 2Central Department of Microbiology, Tribhuvan University, Kirtipur, Nepale-mail: firstname.lastname@example.org AbstractRandom Amplified Polymorphic DNA (RAPD) is a method of producing a genetic fingerprint of a [more...]
Date: 2010-01-21 14:06:00
Cytoplasmic & Nuclear RNA Purification Kit
RT-PCR was performed using human beta actin primers on 2 µL of the 50 µL of cytoplasmic RNA isolated from 1 million HeLa cells using Bio-Synthesis's Cytoplasmic & Nuclear RNA Purification Kit. Lane M is Bio-Synthesis's PCRSizer DNA ladder, Lanes 1 and 3 are the negative control (PCR only, without reverse transcript), and Lanes 2 and 4 are the actual RT-PCR that show the expected 166 bp RT-PCR product. The expected amplicon size from the gene copy is the same as the that from the RNA transcript. The lack of a product in lanes 1 and 3 indicate that no genomic DNA contamination is present in [more...]
Date: 2010-08-16 04:58:00
Basics: Sequencing DNA, Part 1 « Genetic Inference
However, in certain respects Sanger Sequencing is still top-of-the-range. The 4-channel capillary approach lets you sequence DNA pretty fast, and each machine can often do hundreds of reactions at once. It was these machines that first sequenced the human genome. In addition, you can sequence relatively long sequences of DNA; up to 1000 nucleotides (after that, it gets difficult to tell the difference between molecules of different sizes). We have yet to make a machine that can sequence DNA fragments of this length faster than Sanger Sequencing (yet…). The modern Sanger Sequencing machines [more...]
Date: 2009-04-17 21:31:39
Free molecular biology software for Macs | Bioinformatics Blog
Posted on the October 12th, 2010 under Free mac software by bioinformatics-blog 1. Serial Cloner Serial Cloner is fantastic all-in-one workbench; import and manipulate sequences, construct plasmid and restriction site maps, determine %GC and fragment TM, extract and ligate fragments, perform virtual PCR… and lots more, all in one window using a very intuitive graphical interface. 2. 4Peaks 4Peaks is an extremely user friendly DNA sequence chromatogram viewer and editor from the extremely talented guys at Mekentosj. It’s miles better than any of it’s clunky counterparts… try it, [more...]
Date: 2010-10-12 11:08:47
The Quality Stocks Stock Newsletter For Smallcap Companies Blog ...
Shivji noted the 15 years of genomic research and leadership experience of Dr. Schroth, and his tireless efforts to advance gene expression techniques using DNA sequencing, microarrays, PCR and next-gen sequencing, all of which has resulted in numerous publications and patents spanning the entire field of genomic analysis. Dr. Schroth expressed his excitement over the opportunity to push the SmartChip platform forward as a perfect solution for multiple global research efforts that are focused on making improved and more targeted therapeutics, and called the SmartChip system “the most [more...]
Date: 2010-10-12 17:57:42
TriMark Publications Announces Release of Its Nutraceuticals and ...
Packed with the latest information relating to new products and industry trends, this analysis not only quantifies but also qualifies the nutraceuticals market in the areas of research, product development and investment opportunities. ... DNA Sequencing and PCR Markets - Genomics World Markets - High-Growth Diagnostic Tests Markets - Indian Pharmaceutical Industry - Key Diagnostic Testing Markets - Mammography World Markets - Medical Imaging Markets - Microarray Markets [more...]
Date: 2010-10-11 21:42:02
Colony PCR: Another Use For Toothpicks « Promega Connections
Colony PCR: Another Use For Toothpicks July 26, 2010 by Isobel I still remember my JOY (sadly, yes) when I first learned about colony PCR. It allowed me to avoid a laborious procedure involving genomic DNA isolation, restriction digestion, and (the dreaded) Southern blotting. I was trying to show definitively that a transposon had successfully been inserted into a target gene. Using colony PCR, I could just amplify the DNA from a colony and then show that the transposon increased the size of the expected PCR product compared to a control. Joy. I could also use colony PCR to screen DNA [more...]
Date: 2010-07-26 15:13:31
PLoS ONE: Analysis of Short Tandem Repeats by Parallel DNA Threading
Abstract TopThe majority of studies employing short tandem repeats (STRs) require investigation of several of these genetic markers. As such, we demonstrate the feasibility of the trinucleotide threading (TnT) approach for scalable analysis of STRs. The TnT method represents a parallel amplification alternative that addresses the obstacles associated with multiplex PCR. In this study, analysis of the STR fragments was performed with capillary gel electrophoresis; however, it should be possible to combine our approach with the massive 454 sequencing platform to considerably increase the [more...]
Date: 2009-11-13 08:00:00
Gerard Biotech Spin Doctor Genomic DNA isolation kits
**From Tails to PCR in under 2 hours!** +Individual Lypilized tubes can digest samples for PCR in 1-2 hours +15 minute post digest cleanup for +PCR, 30 minute digest for Southerns May be stored safely at room temperature, no need to freeze or aliquot **No column, binding resin, or phenol** +Great yeilds from small samples; 1 mm tail snips, ear punches, young mice +Gentle process will not shear DNA +Eliminates the need for hazardous organic solvents [more...]
Date: 2010-09-29 10:13:26
Extraction of DNA from Fish Fins (Amrita University)
Many different methods and technologies are available for the isolation of genomic DNA. Here, we describe a method for the isolation of PCR-ready genomic DNA from zebra-fish tissues. Video Rating: 0 / 5Source: DNA [more...]
Date: 2010-10-09 19:26:00
Photo Irl | Stadium Sports Central
... features can be built to expand the range of PCR-based applications. PCR: A Practical Approach Volume 2 is not a revised version of PCR: A Practical Approach, but sets out to address some of the new and exciting applications of PCR including direct sequencing, mRNA quantitation and expression, genomic DNA mapping, and mutation analysis. .... Irl Allison Library. Arranged by Irl Allison. This edition: 9569. Willis. Book only. 32 pages. Published by Willis Music. [more...]
Date: 2010-10-12 22:37:02
Epigenetics Papers: A Genomic Sequencing Protocol that Yields a ...
Genomic sequencing protocols, which have been developed to ascertain the methylation status of selected regions within genes, utilize the Maxam and Gilbert chemical cleavage reactions [. ... To address these problems, we have developed a genomic sequencing method that provides positive identification and localization of 5-methylcytosine in genomic DNA. The method is based on sodium bisulfite-mediated conversion of cytosine to uracil in single-stranded DNA, followed by PCR [more...]
Date: 2007-03-13 21:09:00
Nuclear or genomic DNA is found in all nucleated cells as well as in the reproductive cells (eggs and sperm). The amount of DNA we can expect to find in different cells and types of evidence is found in Table 1. ... The second method is a specialized 'nonorganic' extraction using Chelex beads. Chelex beads can only be used when PCR-based DNA testing is going to be used. The basic DNA extraction procedures, whether organic or non-organic, can be adapted for special [more...]
Date: 2010-08-01 06:59:59
Forensic Serology Information Page
What is Forensic Serology?According to forensic serologist Marcella Jones of www.ForensicMentor.com, forensic serology is the analysis of body fluids as they relate to forensic cases, including DNA analysis. Accordingly, the role of the forensic serologist involves:Examining evidence for the presence of body fluids e.g. blood, semen, hair, tissue, saliva, feces, and urineEvaluating evidence for potential DNA analysisEvaluating species of body fluidsPerform PCR (polymerase chain reaction) based typing of STR (short tandem repeat sequences) of genomic DNA, or mitochondrial sequence analysis of [more...]
Date: 2010-02-04 08:00:00
An ELISA-based high throughput protein truncation test for ...
Here, we report the next generation HTS-PTT using triple-epitope-tagged proteins and demonstrate, for the first time, its efficacy on clinical genomic DNA samples for BRCA1/2 analysis. Methods: Segments of exons 11 of BRCA1/2 open reading frames were PCR amplified from either blood derived genomic DNA or cell line mRNA. PCR primers incorporate elements for cell-free transcription/translation and epitope tagging. Cell-free expressed nascent proteins are then antibody-captured onto the wells of a microtiter plate and the relative amount of truncated polypeptide measured using antibodies against [more...]
Date: 2010-10-04 16:47:13
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
Genome Biology | Full text | Optimization and clinical validation ...
Novel features of this platform include the development of a robust algorithm that accurately predicts PCR bias during DNA amplification and can be used to improve PCR primer design, as well as a powerful statistical concept for inferring .... This probability is reduced when intra-primer hairpin formation is predicted, and increased according to degree of complementarity between tag sequence and viral sequence. In this manner, the probability that each nucleotide will be [more...]
Date: 2007-05-27 00:00:00
Evaluation of Five Methods for Total DNA Extraction from Western ...
DNA extraction is a routine step in many insect molecular studies. A variety of methods have been used to isolate DNA molecules from insects, and many commercial kits are available. Extraction methods need to be evaluated for their efficiency, cost, and side effects such as DNA degradation during extraction. Methodology/Principal Findings From individual western corn rootworm beetles, Diabrotica virgifera virgifera, DNA extractions by the SDS method, CTAB method, DNAzol® reagent, Puregene® solutions and DNeasy® column were compared in terms of DNA quantity and quality, cost of materials, [more...]
Date: 2010-08-13 07:00:00
Molecular Clinical Lab Scientist
This achievement was unthinkable only 5 years ago. Be part of a leading edge team working with the best genomic tools. Duties include: Perform all aspects of genetic testing such as DNA extraction, PCR, [more...]
Date: 2010-09-19 23:18:22
In Vivo Assembly of DNA Constructs-Columbia Technology Ventures
Eliminates the use of restriction enzymes and many PCR amplification steps by employing the use of double stranded DNA breaks to trigger in vivo homologous recombination • Equally applicable to small as well as large DNA fragment assembly • The cycling of only two selectable markers and the use orthogonal endonucleases allows for repeated rounds of assembly of many DNA fragments • The assembled DNA is in a known location of the yeast genome and can thus be easily transferred from one strain to another Patent Status: PCT/US10/32962 Licensing Status: Available for Licensing and Sponsored [more...]
Date: 2010-10-02 16:40:33
sequencing of short DNA fragments (40-240bp) - SEQanswers
Hello together, We would like to prepare barcoded fragment libraries of enzymatically digested genomic DNA. The fragment size range we are interested in is 40 to 240bp. We want to sequence 50bp using a SOLiD 4 machine. My questions are: 1) What will happen if e.g. a 40bp fragment is sequenced? Will sequencing continue into the internal adaptor? There are various blocking oligos in the sequencing kit, and I am not entirely sure about their purpose. If they block the complete internal adaptor sequence, does this mean that only 34bp will be sequenced (I believe the fluorescent oligos are [more...]
Date: 2010-08-31 11:57:47
(IUCr) Structure of BT_3984, a member of the SusD/RagB family of ...
The gene encoding BT_3984 (GenBank NP_812895; Swiss-Prot Q8A0N7) was amplified by polymerase chain reaction (PCR) from B. thetaiotaomicron VPI-5482 genomic DNA using PfuTurbo DNA polymerase (Stratagene) and I-PIPE (Insert) primers [more...]
Date: 2010-09-22 07:00:00
(IUCr) Expression, purification and crystallization of Chaetomium ...
The copy number of the gene integration in the genome of Pichia pastoris was determined to be between 7 and 12. PCR amplifications were carried out based on genomic DNA using primers corresponding to [more...]
Date: 2010-08-28 07:00:00
LAB TECH – DUKE – Molecular biology/evolution/genomics (Durham, NC ...
DUTIES: Responsibilities will include: DNA and RNA extractions, sample prep for Illumina library construction and sequencing, standard and quantitative real-time PCR, maintenance of live insects in the lab, as well as lab organization, ordering, and management. Experience with any or all of the above is desirable. The first few weeks of the position will involve helping to set up the lab (ordering, etc). Depending on skills and interests, opportunities could also include genome sequence analysis, use of molecular databases, molecular phylogenetics, design of PCR and sequencing primers, field [more...]
Date: 2010-10-11 18:43:41
Efavirenz Plasma Concentrations and Cytochrome 2B6 Polymorphisms ...
Genomic DNA was extracted from the peripheral whole blood of each subject with a DNA extraction kit (Qiagen, Valencia, CA). CYP2B6 polymorphisms were genotyped by resequencing using primers and polymerase chain reaction (PCR) conditions described by Lamba and associates.15 Sequencing was carried out with an ABI Prism 3700 Automated Sequencer using the PCR primers or internal primers (sequence available on request). Sequences were assembled using the Phred-Phrap-Consed [more...]
Date: 2010-09-22 19:28:38
Epigenetics Papers: DNA Methylation Analysis by MethyLight Technology
Authors discussed each step for this procedure: (1) determining the site of interest for methylation analysis; (2) methylation-specific primers and fluorogenic probes design; (3) genomic DNA isolation; (4) bisulfite conversion; (5) real- time methylation analysis; and (6) data processing. For primer design, some complications particular to the bisulfite modification should be taken into account: All unmethylated cytosine residues in the genome are converted this reduction in genomic complexity reduces the annealing specificity of PCR primers and fluorogenic probes, which not only [more...]
Date: 2007-08-19 01:26:00
Is there a different type of primer i have to use when running PCR ...
Question by bob oso: Is there a different type of primer i have to use when running PCR with genomic DNA compared to cDNA? Im still in school but when i run a PCR, I normally isolate RNA first then RT to cDNA and run PCR with that. This time i isolated the genomic DNA from the sample and ran PCR using the beta-actin primer i always used. Dimers were faintly visible on my gel but nothing really came up like it normally would have or should have. I was told that when running PCR with genomic DNA, a different kind of primer had to be used; one that is specific to genomic DNA. Im confused as to [more...]
Date: 2010-10-01 19:54:46
DNA method gives new perspective on the Mysteries of Nature
We demonstrate that this new approach enables the assignment of virtually all the generated DNA sequences to the correct source once sequencing anomalies are accounted for (miss-assignment rate less than 0.4%). Therefore, the method enables accurate sequencing and assignment of homologous DNA sequences from multiple sources in single high-throughput GS20 run. We observe a bias in the distribution of the differently tagged primers that is dependent on the 5' nucleotide of the tag. In particular, primers 5' labelled with a cytosine are heavily overrepresented among the final sequences, while [more...]
Date: 2007-02-15 17:01:00
Annals of Clinical Microbiology and Antimicrobials | Full text | A ...
© 2010 Williams et al; licensee BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. AbstractBackground Chronic lung infection with the bacterium Pseudomonas aeruginosa is one of the hallmarks of cystic fibrosis (CF) and is associated with worsening lung function, increased hospitalisation and reduced life expectancy. A virulent clonal strain of P. [more...]
Date: 2010-07-16 00:00:00
genomic DNA PCR protocols and methods
Research news, bioproducts, protocols, reagents, forums genomic DNA PCR > Can someone forward me a general protocol for doing PCR using genomic > DNA as the template? I seem to recall needing about 100 ng of DNA, but > I forget stuff like how much Taq to use, concentration of dNTP's, and > concentration of primers. General protocol for PCR Each person uses a slightly different ratio of reagents. For DNA amplicons up to 500 bp, use 25 uL reaction volumes as follows: On ice add to thin-walled pcr tubes 0.125 uL Taq polymerase (1.25 U, Gibco) 2.5 uL of 10 x PCR buffer minus Mg [more...]
Date: 2004-12-27 00:00:00
Heliscope builds world's fastest DNA sequencer | Singularity Hub
The Short: Technology Review reports that for just $1,350,000 USD you can have have the worlds fastest commercially available DNA sequencer, called the Heliscope. The machine developed by Helicos BioSciences takes just one hour to read 1.3 billion base pairs from a strand of DNA. Here is a picture of the beauty: HeliScope™ Single Molecule Sequencer The Long: The Heliscope is being marketed as a DNA microscope. It is unique in the field of DNA sequencing because it sequences an actual DNA strand whereas most other sequencing technologies use PCR to create millions of copies of an original [more...]
Date: 2008-07-14 16:58:33
Eppendorf to market Akonni TruTip™ « Frederick County Biotech ...
HAUPPAUGE, N.Y. & FREDERICK, Md.–(BUSINESS WIRE)–Eppendorf North America and Akonni Biosystems today announced they have entered into a joint marketing agreement to promote Akonni TruTip nucleic acid extraction kits configured for use with Eppendorf epMotion automated pipetting systems. Under the agreement, Eppendorf will promote the extraction kits to clinical, clinical research and forensic laboratories in North America – providing users with access to the industry’s most rapid and reliable means to automatically extract PCR-ready DNA and/or RNA from larger volume [more...]
Date: 2010-04-26 16:36:03
DNA: Size Does Matter. But is it Science or Art? « Exploratorium ...
Then I randomly found this website, which starts with the same idea as our DNA Cheek Cell Extraction Demo but takes it to a whole new level: You send your cheek cells in the mail to their company. They extract your DNA from your cells, then use PCR to amplify unique sections of DNA in your genome. This DNA is loaded into a gel and a current is run through the gel to separate the DNA based on size. UV dye is added to the whole thing to make your DNA strands visible, a photograph is taken, and voila!…a giant canvas print with the color scheme of your choice is ready to hang on your [more...]
Date: 2008-05-14 07:37:47
PLANT TISSUE CULTURE TECHNIQUE AND PCR TECHNOLOGY
SSR assays require a minimal amount of genomic DNA. Most of the crop improvement programs are confined to evaluation and selection of naturally occurring clonal variations. In vitro culture techniques provide an alternative means of plant propagation and an important tool for crop improvement. The in vitro cultures are also of low risk for genetic variation since it is more resistant to genetic changes while occurrence of cell division in vitro condition. Gel electrophoresis is an important molecular biology tool which enables us to study DNA. It can be used to determine the sequence of [more...]
Date: 2010-09-08 04:07:38
Intellectual Property Expert Group (ipeg) » Blog Archive » Are the ...
These are just two recent cases in the US but the implication on DNA sequence patents may be far reaching. Back in 2000, when the new economy was still alive, there was a lot of focus on DNA sequence patents. A lot of companies were very cautious and a lot of genetic test developments were stalled because of patent issues. When asking an expert you could get philosophical answers: either you licence or you ignore or you litigate. Now in 2000 there were still patents from the early days of DNA sequencing around. Some of these patents might have been inventive as the inventive step was a lot [more...]
Date: 2010-09-20 16:55:26
Ganit Labs Bangalore Genomics/DNA Microarray Scientist/RA/Lab ...
Ganit Labs Bellary Road, Hebbal, Bangalore 560024 Positions: Scientist/RA/Lab Director/Informatics Manager Genomics Applications and Informatics Technology Labs (Ganit Labs) is a newly formed, Bangalore-based, public private-partnership initiative. Ganit Labs would work in the field of functional genomics and next generation sequencing. Currently we are building a state-of-the art modern genomics lab along with the best computational facility that would present entrepreneurial atmosphere of start-up companies and academic rigor and solid training of research institutes/universities. The [more...]
Date: 2010-08-22 04:26:12
Variants of PCR (1)
The bacteriophage T4 DNA polymerase was also tested shortly after the first reports of PCR. It has a higher fidelity of replication than the Klenow fragment. Since it is also destroyed by heat, it has seen little use since the development of thermostable polymerases. The DNA polymerase from Thermus aquaticus (or Taq), was the first thermostable polymerase used in PCR, and is still the one most commonly used. The enzyme can be isolated from its 'native' bacterial source, or from a cloned gene expressed in E. coli. The Stoffel fragment is produced from a truncated gene for Taq polymerase, [more...]
Date: 2008-09-11 03:56:00
Garage biotech in Nature magazine | All my RSS
Want to know more about the DIYbio scene? This month's Nature magazine features an excellent piece covering garage biotechnology. For now, most members of the do-it-yourself, or DIY, biology community are hobbyists, rigging up cheap equipment and tackling projects that -- although not exactly pushing the boundaries of molecular biology -- are creative proof of the hacker principle. Meredith Patterson, a computer programmer based in San Francisco, California, whom some call the 'doyenne of DIYbio', made glow-in-the-dark yogurt by engineering the bacteria within to produce a fluorescent [more...]
Date: 2010-10-12 00:00:00
Genetic Fingerprinting of Bacillus thuringiensis Isolates by ...
RAPD-PCR mixture was set up that contained 20-50 ng of genomic DNA, 40 pmol of primer, 1 U of Taq polymerase (Bangalore GENEI.), 200 uM (each) deoxynucleoside triphosphate, 10 mM Tris-Cl (pH 8.3), 50 mM KCl, 3 mM MgCl2 and 1% DMSO. Each reaction mixture was overlaid with 25 ul of mineral oil and amplified with a Perkin-Elmer Cetus DNA Thermal Cycler model TC-1 as follows: (i) Initial denaturation, 1 cycle consisting of 5 min at 94oC and (ii) 35 cycles, with 1 cycle consisting of 1 min at 94oC, 2 min at 36oC, and 3 min at 72oC, followed by a final extension step at 72oC for 10 min. RAPD [more...]
Date: 2009-09-25 16:32:00
Research Technician vacancy in Biology at Univ Hospital Zurich ...
Applicants should hold either a degree as professional technician (e.g. SRK), or an MSc in Biology, and must have a strong practical background in the following technologies: genomic DNA extraction/purification, quantitative PCR and good computer knowledge. Experience with robotic equipment and automation are an advantage. Skills in laboratory management, high level of self-organization and interpersonal skills to work within an international group of scientists are a plus. Fluent technical English and German are obligatory. Applicants should submit a letter of interest, CV, working [more...]
Date: 2010-10-11 22:47:53
GENOMIC CHARACTRIZATION OF AEROMONAS HYDROPHILA ISOLATES USING ...
GENOMIC CHARACTRIZATION OF AEROMONAS HYDROPHILA ISOLATES USING RAPD-PCR TECHNIQUE by Mitali Dhiman and S. S. Mishra Central Inland Fisheries Resear Aeromonas hydrophila are involved in various disease problems in humans and aquatic animals and are known to be phenotypically, serologically and genetically quite diverse. Development and use of a sensitive and specific diagnostic test is warranted for detection and characterization of this pathogen.In the present study, Fish and water samples from river Hooghly were streaked onto Aeromonas selective growth medium (Rimler-shotts [more...]
Date: 2010-10-08 20:08:05