Navigation Links
Preparation of PCR Quality Template DNA from a Wide Range of Tissues Using InstaGene Matrix

Use 20 l of the supernatant for each PCR reaction.

Notes: Store remainder at -20 C for subsequent PCRs. Repeat step 6 when reusing the preparation (allow to thaw first).


PCR
Primers used in this experiment were designed to amplify a 120 base pair fragment of the Canine Pancreatic Lipase Gene (Mickel et al., 1989). Amplification was carried out using 2 Units AmpliTaq DNA polymerase (Perkin-Elmer) mixed with an equal volume of TaqStart antibody (Clontech) per reaction in a total reaction volume of 50 l, containing 50 pmol each primer and 200 mol/L of each deoxynucleoside triphosphate in 10 mM Tris (pH 8.4), 50 mM KCl, and 1.5 mM MgCl2. Thermal cycling was performed on a Perkin-Elmer 9600. Initial denaturation was at 94 C for 5 minutes, followed by 40 cycles of 94 C for 30 seconds, 57.5 C for 1 minute, then a final extension of 72 C for 7 minutes.


Results

All tissues examined produced a clear band of the correct size. The variation in the brightness of the band does not appear to correspond to the amount of DNA template present in each reaction. Apart from the positive control (A72 cell line), there are two template copies per cell for each tissue. The most likely cause of the variation in intensity of the band obtained is therefore inhibitory factors present within the sample. It is important not to exceed the capacity of the InstaGene matrix by using too much tissue in the initial preparation. Exceeding the matrix capacity may cause PCR-inhibitory factors normally removed by the procedure to remain in the supernatant. Carryover of these inhibitory factors may affect amplification in the subsequent PCR reaction.
'"/>

Source:


Page: All 1 2 3

Related biology technology :

1. Sample Preparation for Microinjection
2. High Pure PCR Template Preparation Kit
3. Proteinase K, Recombinant, PCR Grade: the Ideal Tool for Template Preparations
4. Dicer vs. RNase III for Preparation of siRNA Cocktails
5. Genomic DNA Preparation from RNAlater Preserved Tissues
6. Commercial Plasmid Preparation Methods Yield DNA With Different Transfection Capabilities
7. Sample Preparation Solutions for Cellular Protein Fractionation Poster, Rev A
8. High Throughput Plasmid DNA Preparation Using A Magnetic Bead Kit On An Automated Workstation
9. A Simple and Rapid Technique to Purify High-Quality Midiprep DNA
10. Generate High-Quality, Directional Plasmid cDNA Libraries
11. Quality control of antibodies using the 2100 bioanalyzer and the Protein 200 Plus assay
Post Your Comments:
(Date:8/18/2014)... Global HIV Infection Drug Market and ... Reportstack. The rate of HIV infection and the number of ... at a positive rate across the globe. It has been ... 2005 to 1.5 in 2013, with the rates for children ... changes in the infection rates has been the improved access ...
(Date:8/18/2014)... 2014 According to the ... Application (Municipal Wastewater Treatment and Industrial Wastewater Treatment), ... Tubular), By Configuration (Internal/Submerged and External/Sidestream), and By ... The market for membrane bioreactor systems is projected ... a CAGR of 15.28% between 2014 and 2019. ...
(Date:8/18/2014)... JUPITER, Fla. , Aug. 18, 2014 /PRNewswire/ ... global biotechnology company whose patented and proprietary technologies ... and other proteins for the bioenergy, bio-based chemical, ... it has filed a Registration Statement on Form ... (the "SEC"). Logo - http://photos.prnewswire.com/prnh/20110621/CL06708LOGO ...
(Date:8/18/2014)... August 2014 Vier führende deutsche ... klinische Studie auf, in der eine Intraokularlinse ... das Sehvermögen bei allen Entfernungen verbessern, so ... keine Lese- oder Gleitsichtbrille mehr benötigen. Die ... FluidVision Accommodating Intraocular Lens (FluidVision akkomodative ...
Breaking Biology Technology:Global HIV Infection Drug Market Overview 2Global HIV Infection Drug Market Overview 3Global HIV Infection Drug Market Overview 4Membrane Bioreactor Systems Market Worth $2,927 Million by 2019 2Membrane Bioreactor Systems Market Worth $2,927 Million by 2019 3Dyadic International Files Form 10 Registration Statement 2Dyadic International Files Form 10 Registration Statement 3Dyadic International Files Form 10 Registration Statement 4Implantation einer revolutionären akkommodativen Linse bei Katarakt-Patienten in deutschen Augenzentren im Rahmen einer internationalen klinischen Studie 2Implantation einer revolutionären akkommodativen Linse bei Katarakt-Patienten in deutschen Augenzentren im Rahmen einer internationalen klinischen Studie 3Implantation einer revolutionären akkommodativen Linse bei Katarakt-Patienten in deutschen Augenzentren im Rahmen einer internationalen klinischen Studie 4
... heat during the production of electricity could be ... via a technique developed by researchers with the ... Laboratory (Berkeley Lab) and the University of California ... this technology include DOEs hydrogen fuel cell-powered Freedom ...
... in the Philadelphia suburbs, with unique new capabilities enabled by Masimo ... ... technologies, IRVINE, Calif., Jan. 10 Abington Memorial,Hospital and ... & Low Perfusion Pulse Oximetry, have announced the,completion of Philadelphia-based Abington ...
... Ky., Lexington, Ky., and Newport News, Va. Also ... ... spirit of excellence and remarkable achievements in its,community, Harborview Medical Center of ... is one of the most esteemed,honors of excellence in community service in ...
Cached Biology Technology:Feeling the heat 2Feeling the heat 3Feeling the heat 4Abington Memorial Hospital Completes System-Wide Conversion to Masimo SET Pulse Oximetry Technology 2Abington Memorial Hospital Completes System-Wide Conversion to Masimo SET Pulse Oximetry Technology 3Abington Memorial Hospital Completes System-Wide Conversion to Masimo SET Pulse Oximetry Technology 4Harborview Medical Center in Seattle Receives Prestigious $100,000 Foster G. McGaw Prize for Excellence in Community Service 2Harborview Medical Center in Seattle Receives Prestigious $100,000 Foster G. McGaw Prize for Excellence in Community Service 3Harborview Medical Center in Seattle Receives Prestigious $100,000 Foster G. McGaw Prize for Excellence in Community Service 4Harborview Medical Center in Seattle Receives Prestigious $100,000 Foster G. McGaw Prize for Excellence in Community Service 5Harborview Medical Center in Seattle Receives Prestigious $100,000 Foster G. McGaw Prize for Excellence in Community Service 6
(Date:8/18/2014)... Half of the Earth,s land mass is made ... they are being transformed at an alarming rate. Woody ... and taking over, leading to a loss of critical ... of ecosystems to produce food specifically meat. ... Sciences led an investigation that quantified this loss in ...
(Date:8/18/2014)... Researchers from Aarhus University, Denmark, and the University of Wisconsin-Madison, ... exposure of species to both climate and land use change. ... species in the face of combined rates of climate and ... , Their results, which have just been published in ... vulnerable to losses in biodiversity and ecosystem function due to ...
(Date:8/18/2014)... the pygmy phenotype is probably a selective adaptation ... of researchers, but all African pygmy phenotypes do ... more recent adaptation than previously thought., "I,m interested ... very challenging environments," said George H. Perry, assistant ... rainforests are difficult for humans to live in. ...
Breaking Biology News(10 mins):Trees and shrubs invading critical grasslands, diminish cattle production 2Trees and shrubs invading critical grasslands, diminish cattle production 3The double threat of climate and land use change enhances risks to biodiversity 2Pygmy phenotype developed many times, adaptive to rainforest 2
... 28, 2007 - Sirtris Pharmaceuticals, Inc. (NASDAQ: SIRT), a ... drugs to treat diseases of aging, announced today that ... developed novel drug candidates that offer a promising, new ... Diabetes, by targeting SIRT1, a gene that controls the ...
... Although a high standard of morality gives but a ... his children over the other men of the same ... give an immense advantage to one tribe over another. ... an evolutionary explanation for morality and pro-social behaviors individuals ...
... able to form abstract concepts. Friederike Range and ... shown for the first time that dogs can classify ... the same way that humans do. And the ... computer automated touch-screens, eliminating potential human influence. The ...
Cached Biology News:Sirtris unveils promising, novel SIRT1 activators for treating diseases of aging 2Sirtris unveils promising, novel SIRT1 activators for treating diseases of aging 3Group selection, a theory whose time has come...again 2Computer savvy canines 2
Gloves are 9.25 long, latex, powder free, smooth and ambidextrous....
Request Info...
Prepared with distilled water. Yeast extract solution is autolyzed at 150 g/L....
Streptomycin sulfate an aminoglycoside used for bacterial selection binds to the small ribosomal subunit and interferes with protein biosynthesis.21H39N7O12)2x3H2SO4Formula weight:...
Biology Products: