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:9/30/2014)... Using a bio-mimicking analog of one of ... an international research team led by Alejandro Briseno ... a major step in developing long-sought polymer architecture ... for use in electronic devices. , Briseno, ... and others at Stanford University and Dresden University ...
(Date:9/30/2014)... 30, 2014 Nematicide is a type ... are microscopic parasitic roundworms, found in massive quantities ... plants and animals. Nematicides have tended to be ... promoting migration through the soil. A single gram ... nematodes. With more than 10,000 species classified, they ...
(Date:9/30/2014)... Sept. 30, 2014  RegeneRx Biopharmaceuticals, Inc. (OTC: ... it has received a Canadian Patent for Inhibition ... This patent includes claims using thymosin beta 4, ... peptides for these purposes.  The patent will expire ... Inc. ( www.regenerx.com ) RegeneRx ...
(Date:9/30/2014)... Angeles, CA (PRWEB) September 30, 2014 ... (EWS), the high-speed, chemical-free process to clean up large ... to shareholders, CEO Riggs Eckelberry reported on the launch ... Texas, at a site provided by partner STW ... up to 1000 barrels per day (bpd) and achieved ...
Breaking Biology Technology:Blades of grass inspire advance in organic solar cells 2Blades of grass inspire advance in organic solar cells 3Nematicides Market by Types (Fumigants, Organophosphates, Carbamate and Bio-Nematicides), Applications (Agrochemicals, Industrial, and Others), and Geography (North America, Europe, Asia-Pacific & ROW) - Global Trends & Forecasts to 2019 2RegeneRx Receives Canadian Patent for Inhibition or Reversal of Skin Aging 2OriginOil, Inc. Report to Shareholders: Blue Gold in West Texas 2
... Biotechnologies AB, an,atherosclerosis R&D company developing novel products ... today that it has,signed two distributor agreements - ... Ltd. , Carina Schmidt, ... about the,opportunities these agreements present to Athera. They ...
... that Understanding the Fundamental Mechanism of Angiogenesis is ... - AAmgen Launches Angiogenesis Website THOUSAND OAKS, Calif., ... ) today announced that results from several preclinical ... presented at the 2009 American Association for Cancer ...
... SAN FRANCISCO, Calif., April 16 Proteolix, Inc. today ... post of Senior Vice President, Finance and Corporate Development ... the company,s clinical development and regulatory leadership team with ... the post of Chief Medical Officer (CMO) and Albert ...
Cached Biology Technology:Athera Signs Agreements With German and UK Distributors for New Cardiovascular Biomarker 2Athera Signs Agreements With German and UK Distributors for New Cardiovascular Biomarker 3Preclinical Data From Amgen's Oncology Pipeline to be Presented at AACR 2Preclinical Data From Amgen's Oncology Pipeline to be Presented at AACR 3Preclinical Data From Amgen's Oncology Pipeline to be Presented at AACR 4Preclinical Data From Amgen's Oncology Pipeline to be Presented at AACR 5Preclinical Data From Amgen's Oncology Pipeline to be Presented at AACR 6Preclinical Data From Amgen's Oncology Pipeline to be Presented at AACR 7Proteolix Announces New Appointment in Finance and Expands Clinical and Regulatory Leadership Team 2Proteolix Announces New Appointment in Finance and Expands Clinical and Regulatory Leadership Team 3Proteolix Announces New Appointment in Finance and Expands Clinical and Regulatory Leadership Team 4
(Date:9/30/2014)... Calif. The National Institutes of Health (NIH) ... today to develop an electrode array system that ... brain works through unprecedented resolution and scale. , ... to support President Obama,s BRAIN (Brain Research ... research effort to revolutionize our understanding of the ...
(Date:9/30/2014)... live in almost any aquatic environment on Earth, but ... species are pushed to the limit. The amount of ... critical for how different species cope in the future, ... of Gothenburg, published in the scientific journal Proceedings ... continues apace thanks to increasing levels of greenhouse gases ...
(Date:9/30/2014)... want your doctor to know what goes wrong with your ... idea to know what "normal" actually is. That,s where a ... the FASEB Journal comes in. In ... key set of molecules that can help scientists "see" which ... What,s more, they found never-before-detected gene activity and that men ...
Breaking Biology News(10 mins):NIH taps lab to develop sophisticated electrode array system to monitor brain act 2NIH taps lab to develop sophisticated electrode array system to monitor brain act 3Shape up quickly -- applies to fish, too! 2Scientists identify which genes are active in muscles of men and women 2
... paleobiologist J. William Schopf and colleagues have produced 3-D images ... old -- preserved in rocks, an achievement that has never ... Mars brings rocks back to Earth, Schopf said the techniques ... spectroscopy, could enable scientists to look at microscopic fossils inside ...
... National Jewish Medical and Research Center report in the ... promising proteins, known as fibulins 3 and 5, slow ... blood vessels from sprouting. The proteins are promising candidates ... produce fibulin proteins, which regulate cell proliferation, migration and ...
... be potentially life threatening in the home, but the presence ... Mike Steven and colleagues from the University of Nottingham have ... act as an early warning of leaks in natural gas ... satellite remote systems to monitor gas leaks via the spectrum ...
Cached Biology News:Inside rocks, implications for finding life on Mars 2Proteins stop blood-vessel and tumor growth in mice 2Plants used to detect gas leaks, from outer space! 2
... This CLS number ... number, created to easily ... If showing no availability ... the old Sigma-Aldrich number ...
This CLS number is a new product number, created to easily match Cornings product number. If showing no availability yet, please order under the old Sigma-Aldrich number (E6027) or contact customer s...
... pressure gradient system uses three Adept CE ... a flow rate in one microlitre increaments. ... added flexibility. System control and data processing ... interface unit, CE 4900., Column heater/chillers can be ...
... of large numbers of samples may be carried ... minimum operator attention. System control and data ... CE 4900. The elegant, powerful and very ... and Windows 98 and 2000., The supplied detector ...
Biology Products: