Navigation Links
The Use of MasterTaq Polymerase for PCR with Humic Material-Contaminated ,,, DNA

Katrin Ravenschlag
Max-Planck-Institute for Marine Microbiology, Celsiusstr.1, 28359 Germany
Phone: ++49-421-2028-936, Fax: ++49-421-2028-580, email: kravensc@mpi-bremen.de

Introduction

Direct extraction of total DNA from soils or sediments always results in coextraction of other soil components, mainly humic acids or other humic substances, which negatively interfere with DNA detection and biochemical reactions.1 This contamination can, for example, inhibit Taq Polymerase in PCR*,25 or restriction endonucleases.6,3 Humic material, even in quantities as small as 1 ng, has been shown to inhibit PCR.7 Humic acids and nucleic acids share similar physico-chemical properties, making the complete removal of humic material difficult. Numerous studies on the development of an effective purification method have been conducted but none appears universal (see References 810). In this article, I shall be reporting on the excellent suitability of MasterTaq polymerase for the amplification of 16S rDNA genes from humic substances-contaminated DNA extracted from a marine sediment (Svalbard, Arctic Ocean).

Materials and Methods

DNA extraction. Total DNA was directly extracted from the sediment according to Zhou et al.10 The protocol encompassed three cycles of freezing and thawing, chemical lysis in a high-salt extraction buffer by heating the suspension in the presence of sodium dodecyl sulfate (SDS) and hexadecyltrimethylammonium bromide (CTAB), and a proteinase K step. The crude DNA was purified by dialysis. However, there were humic substances left in the DNA extract indicated by the brownish color of the DNA.


'"/>

Source:


Page: All 1 2 3 4 5

Related biology technology :

1. Amplification of the amoA Gene of Nitrosococcus oceani Bacteria Using Eppendorf MasterTaq
2. Amplification of Mouse cDNAs for Microarrays Using the Eppendorf MasterTaq Kit
3. Enhancing PCR* Efficiency with MasterTaq
4. Novel PCR Enhancing Factor Improves Performance of Pfu DNA Polymerase
5. Pfu DNA Polymerase as a Hot Start Alternative
6. PCR Performance Comparisons Between pfuturbo and Taq DNA Polymerases
7. pfuturbo DNA Polymerase: A High-Performance, High-Fidelity Enzyme Ideal for PCR Cloning
8. Unique DNA Polymerase Formulation Excels in a Broad Range of PCR Applications
9. Enhanced Amplification of Long Targets with PfuTurbo DNA Polymerase
10. High-Fidelity PCR with a Novel Polymerase Mixture
11. Optimizing pfuturbo DNA Polymerase Amplification Reactions with Perfect Match PCR Enhancer
Post Your Comments:
(Date:1/15/2014)... January 15, 2014 This webinar ... on nonclinical and clinical safety assessment in biosimilars. ... guide a path for biosimilar drug development, however ... of biosimilarity with regards to quality, safety and ... of biosimilar drug development and registration, development strategies ...
(Date:1/15/2014)... NC (PRWEB) January 15, 2014 ... an enhancement to its Online Web Portal for Life ... now have the ability to specify the subject matter ... nearly 50 life science sub-domains. This will help reduce ... and scientific fields, and decrease the likelihood of error. ...
(Date:1/14/2014)... Doylestown, PA (PRWEB) January 14, 2014 ... – 11 p.m. , Location: Warrington Country Club, 1360 ... Hepatitis B Foundation, the only national nonprofit organization solely ... improving the quality of life for those affected worldwide, ... 11 at Warrington Country Club in Warrington, Pa. ...
(Date:1/14/2014)... York, NY (PRWEB) January 14, 2014 ... initiated coverage of Alliqua, Inc. (OTCQB: ALQA). Alliqua ... and marketing proprietary products to serve the wound ... , In late 2012, Alliqua was restructured ... launched the company’s new strategy to become a ...
Breaking Biology Technology:Xtalks Life Sciences Webinar Examines Safety Assessment of Biosimilars 2DTS Improves Efficiency for Life Science Document Translations 2Hepatitis B Foundation to Host Annual Crystal Ball Gala 2EquitiesIQ Initiates Coverage of Alliqua, Inc. 2EquitiesIQ Initiates Coverage of Alliqua, Inc. 3
... 3, 2011 EMG Laboratory ... awarded the American Association of Neuromuscular & Electrodiagnostic Medicine,s ... Exemplary Status. EMG Laboratory has provided advanced testing ... around Merrimack Valley and Cape Cod since 1997. Currently, ...
... 2011 Omeros Corporation (NASDAQ: OMER ), ... products focused on inflammation, bleeding and disorders of the ... M.D., chairman and chief executive officer, is scheduled to ... 2011 Canaccord Genuity Growth Conference August 10, 2011 ...
... Inc. (NYSE Amex: CUR ) announced it ... develop its human neural stem cell technology for the ... "Research to Treat Cancerous Brain Tumors with Neural Stem ... Investigator John Zhang, MD, PhD, Professor of Neurosurgery, Loma ...
Cached Biology Technology:EMG Laboratory Attains Prestigious National Accreditation for Clinical Excellence in Neuromuscular Diagnosis 2EMG Laboratory Attains Prestigious National Accreditation for Clinical Excellence in Neuromuscular Diagnosis 3Neuralstem Awarded Department of Defense Contract for Brain Cancer Research 2Neuralstem Awarded Department of Defense Contract for Brain Cancer Research 3
(Date:4/15/2014)... partially funded by the U.S. Department of Agriculture and ... their diets simply by enjoying white or brown rice ... , In a study published online in the peer-reviewed ... Theresa Nicklas, DrPH, of Baylor College of Medicine, analyzed ... 2005-2010 and evaluated the association of rice consumption with ...
(Date:4/14/2014)... you,re over 24 years of age you,ve already reached ... according to a new Simon Fraser University study. ... professor Mark Blair, Thompson,s thesis supervisor, and Andrew Henrey, ... news in a just-published PLOS ONE Journal ... science experiments to rest on big data, the trio ...
(Date:4/14/2014)... levels of the greenhouse gas methane were found above ... to be an important emissions source, according to a ... findings could have implications for the evaluation of the ... which is one of only a few to use ... levels in the air above wells, identified seven individual ...
Breaking Biology News(10 mins):Eating rice boosts diet quality, reduces body weight and improves markers for health 2Eating rice boosts diet quality, reduces body weight and improves markers for health 3Study says we're over the hill at 24 2'Problem wells' source of greenhouse gas at unexpected stage of natural gas production 2
... of cholera appear to have evolved a distinctly different mechanism ... the current issue of the online journal mBio . ... the diarrheal disease cholera. While there are more than ... have been known to cause epidemic and pandemic outbreaks of ...
... 2010) -- When their razor-sharp mandibles wear out, leaf-cutter ... efficient sisters take over cutting, say researchers from two ... of regular publication in the journal Behavioral Ecology ... way of providing for its displaced workers. ...
... endogenous estrogen (i.e., estrogen produced by ovaries and by ... replacement therapy (HRT) utilizing estrogen alone (the exogenous estrogen) ... according to study results presented at the 33rd Annual ... "Our analysis suggests that, contrary to previous ...
Cached Biology News:When their tools get dull, leaf-cutters switch jobs 2When their tools get dull, leaf-cutters switch jobs 3Estrogen alone is effective for reducing breast cancer risk 2
... purification systems provide high speed purification of ... offer increased throughput and reproducible results. The ... mL and KingFisher 96, covering sample volumes ... consists of an instrument, specially designed plastics ...
... The DNA120OP is specifically engineered for efficient ... design incorporates a glass cover and a ... automated, unattended, odor-free processing of samples in ... for Synthetic Oligos ...
... basic SpeedVac concentrator designed for the price-conscious ... still features a standard Teflon coated chamber ... and heat. It is designed for use ... variety of different rotors for maximum sample ...
... SpeedVac to have active, real-time sample temperature ... is the worlds first, fully modular SpeedVac ... stacked together to form an integrated unit ... is also the first SpeedVac to have ...
Biology Products: