Patente US5538871 - In situ polymerase chain reaction - Google Patentes
Improvements to the in situ polymerase chain reaction (PCR), a process of in vitro enzymatic amplification of specific nucleic acid sequences within the cells where they originate, can be achieved by changing the way that the enzymatic reaction is started. Reaction initiation is delayed until the start of PCR thermal cycling, either by withholding a subset of PCR reagents from the cellular preparation until the preparation has been heated to 50 C. to 80 C., immediately before thermal cycling is begun, or by adding to the PCR reagents a single-stranded DNA binding protein which blocks reaction at temperatures below about 50 C. If the in situ PCR is performed on cellular preparations already attached to a microscope slide, thermal cycling also is facilitated by use of a thermal cycler sample block or compartment designed optimally to hold the microscope slide and any vapor barrier covering the slide.http://www.google.es/patents/US5538871
Touchdown polymerase chain reaction - Wikipedia
The touchdown polymerase chain reaction or touchdown style polymerase chain reaction is a method of polymerase chain reaction by which primers avoid amplifying nonspecific sequences. The annealing temperature during a polymerase chain reaction determines the specificity of primer annealing. The melting point of the primer sets the upper limit on annealing temperature. At temperatures just above this point, only very specific base pairing between the primer and the template will occur. At lower temperatures, the primers bind less specifically. Nonspecific primer binding obscures polymerase chain reaction results, as the nonspecific sequences to which primers anneal in early steps of amplification will "swamp out" any specific sequences because of the ...https://en.wikipedia.org/wiki/Touchdown_polymerase_chain_reaction
Mutations of p53 gene can be detected in the plasma of patients with large bowel carcinoma. | Journal of Clinical Pathology
AIMS: To attempt to detect p53 gene mutations in the plasma of patients with large bowel carcinoma. METHODS: Plasma was collected from 20 control patients with no history of cancer and from 17 patients with large bowel carcinoma. Corresponding tumour and benign lymph node (control) samples for each of the carcinoma patients were obtained from paraffin blocks. A Dukes' stage was determined for each tumour. DNA was extracted from the plasma samples and the paraffin embedded tissue using previously described methods. A nested primer polymerase chain reaction protocol was used for the amplification of exons 5 to 8 of the p53 gene. "Cold" single strand conformational polymorphism (SSCP) was performed on mini gels and silver stained. Abnormal bands were excised, the DNA eluted, and reamplified for automated dye termination sequencing. Any sample showing an apparent mutation was rechecked from the original extracted DNA sample at least three times. RESULTS: p53 ...http://jcp.bmj.com/content/51/8/611
Thermal cycler - Wikipedia
The thermal cycler (also known as a thermocycler, PCR machine or DNA amplifier) is a laboratory apparatus most commonly used to amplify segments of DNA via the polymerase chain reaction (PCR). Thermal cyclers may also be used in laboratories to facilitate other temperature-sensitive reactions, including restriction enzyme digestion or rapid diagnostics. The device has a thermal block with holes where tubes holding the reaction mixtures can be inserted. The cycler then raises and lowers the temperature of the block in discrete, pre-programmed steps. The earliest thermal cyclers were designed for use with the Klenow fragment of DNA polymerase I. Since this enzyme is destroyed during each heating step of the amplification process, new enzyme had to be added every cycle. This led to a cumbersome machine based on an automated pipettor, with open reaction tubes. Later, the PCR process was adapted ...https://en.wikipedia.org/wiki/Thermal_cycler
Reference genes for quantitative real-time polymerase chain reaction studies in soybean plants under hypoxic conditions
Quantitative real-time polymerase chain reaction (RT-qPCR) is a powerful tool used to measure gene expression. However, because of its high sensitivity, the method is strongly influenced by the quality and concentration of the template cDNA and by the amplification efficiency. Relative quantification is an effective strategy for correcting random and systematic errors by using the expression level of reference gene(s) to normalize the expression level of the genes of interest. To identify soybean reference genes for use in studies of flooding stress, we compared 5 candidate reference genes (CRGs) with the NormFinder and GeNorm programs to select the best internal control. The expression stability of the CRGs was evaluated in root tissues from soybean plants subjected to hypoxic conditions. Elongation factor 1-beta and actin-11 were identified as the most appropriate genes for RT-qPCR normalization by both the NormFinder and GeNorm analyses. The expression ...http://www.locus.ufv.br/handle/123456789/12041
HKU Scholars Hub: Development and evaluation of a novel in-house single-tube nested real-time polymerase chain reaction assay...
FilmArray is a rapid diagnostic technology which can simultaneously detect multiple pathogens in a single run. It is especially useful in the diagnosis of respiratory diseases because most of the causative pathogens cause similar symptoms. While FilmArray has numerous undisputable advantages in the detection of respiratory pathogens, one of the major concerns is that it has poor sensitivity for adenovirus detection. Reports are consistently showing that the FilmArray Respiratory Panel has only around 50% to 60% sensitivity for the detection of adenoviruses. FilmArray Respiratory Panel's poor sensitivity for adenovirus detection is of particular concern for immunocompromised patients because they are at higher risk of developing severe infection. Therefore, a complementary test would be needed to overcome these limitations of FilmArray. In this study, a novel in-house single-tube nested real-time polymerase chain reaction assay was developed and we evaluated ...http://hub.hku.hk/handle/10722/237229
Polymerase chain reaction-based detection of Mycobacterium tuberculosis DNA in formalin-fixed, paraffin-embedded skin tissue...
Skin and Cancer Foundation, Inc., Pasig City, Philippines, and the National Institute of Molecular Biology and Biotechnology, College of Science, University of the Philippines, ...http://onlinelibrary.wiley.com/doi/10.1111/j.1365-4632.2010.04189.x/full
Inverse polymerase chain reaction - Wikipedia
Inverse polymerase chain reaction (Inverse PCR) is a variant of the polymerase chain reaction that is used to amplify DNA with only one known sequence. One limitation of conventional PCR is that it requires primers complementary to both termini of the target DNA, but this method allows PCR to be carried out even if only one sequence is available from which primers may be designed. Inverse PCR is especially useful for the determination of insert locations. For example, various retroviruses and transposons randomly integrate into genomic DNA. To identify the sites where they have entered, the known, "internal" viral or transposon sequences can be used to design primers that will amplify a small portion of the flanking, "external" genomic DNA. The amplified product can then be sequenced and compared with DNA databases to locate the sequence which has been disrupted. The inverse PCR method involves a series of ...https://en.wikipedia.org/wiki/Inverse_polymerase_chain_reaction
Quantification of the common deletion in human testicular mitochondrial DNA by competitive PCR assay using a chimaeric...
The 'common' 4977 by deletion in mitochondrial DNA (Δ4977) is commonly used as an indicator of tissue deterioration in ageing and bioenergetic diseases. Deletion levels are normally measured by a serial dilution polymerase chain reaction (PCR) approach, where test reactions are compared with dilutions of control amplifications of DNA from a similar sized stable region of the mitochondrial genome. The end-point of this assay is the dilution that can just detect any PCR product; however, this is an inherently unstable measure. We constructed a chimaeric DNA construct that binds to both control and deletion primers with similar annealing properties. This was used in a competitive PCR assay to quantify Δ4977 in human testicular tissues that had been well-characterized using the serial dilution approach. We found the competitive assay to be highly replicable as it compares the PCR product of the construct with that of test DNA samples during the ...http://researchrepository.murdoch.edu.au/id/eprint/17179/
"Development of a Real-Time Polymerase Chain Reaction Detection Method " by C. J. Fitzsimmons, L. Marklund et al.
Myeloperoxidase (MPO) is a hemoprotein present in azurophilic granules of polymorphonuclear (PMN) leukocytes and monocytes. It catalyzes the oxidation of halide ions to their respective hypohalous acids, which are used for microbial killing by phagocytic cells. Measurement of MPO activity is often used as a marker of neutrophil infiltration into tissues. We have designed a quantitative reverse transcription-polymerase chain reaction detection method for porcine MPO transcripts by using TaqMan realtime PCR technology. Total RNA was isolated from lung and spleen tissue collected 7 days post-intranasal inoculation with Salmonella choleraesuis (n=4) or saline (n=4). The lung and spleen samples were pooled before RNA isolation to create uninfected control and infected RNAs for each tissue. Expression of MPO mRNA was highest in infected spleen (12.5 ± 2.85 relative units (RU)), followed by the uninfected spleen sample (2.9 ± 1.50 RU). There was no difference in ...http://lib.dr.iastate.edu/swinereports_2000/20/
A semi-quantitative reverse transcriptase polymerase chain reaction method for measurement of mRNA for TNF-alpha and IL-1beta...
A semi-quantitative reverse transcriptase polymerase chain reaction method for measurement of mRNA for TNF-alpha and IL-1beta in whole blood cultures: its application in typhoid fever and exentric ...http://repository.ubn.ru.nl/handle/2066/23983
Magnetic Beads Genomic DNA Extraction Kit (Blood) MB048/MB096 | Geneaid
The Magnetic Beads Genomic DNA Extraction Kit Blood was designed specifically for efficient genomic DNA purification from blood and buffy coat. DNA is bound to the surface of the magnetic beads and released using a proprietary buffer system.http://www.geneaid.com/products/target-sample/blood/magnetic-beads-blood-dna-extraction-kit
Detection of human papillomavirus type 16 DNA sequences in archival cervical tissues by the polymerase chain reaction. -...
We have evaluated the polymerase chain reaction for the detection of viral DNA sequences in paraffin-embedded archival tissues. In 63 frozen cervical biopsy specimens that were taken from premalignant and invasive lesions, Southern blotting detected human papillomavirus (HPV) type 16 DNA in 28 (44%) of the samples. In the polymerase chain reaction analysis of the formalin-fixed, paraffin-embedded mirror biopsy specimens, 46 (73%) of the tissues were found to be positive for HPV type 16. In three Southern blotting-positive cases, the DNA of the paraffin-embedded sections was too scant or too degraded to allow the detection of HPV DNA by the polymerase chain reaction. In 21 Southern blotting-negative cases, HPV type 16 DNA could be demonstrated in the archival sections by the polymerase chain reaction ...https://www.ndcn.ox.ac.uk/publications/68345
Journal of Current Medical Research and Practice - Real-time quantitative polymerase chain reaction detection of minimal...
This is a temporary file and hence do not link it from a website, instead link the URL of this page if you wish to link the PDF file ...http://jcmrp.eg.net/downloadpdf.asp?issn=2357-0121
신개념 Genomic DNA 추출 키트- MagListoTM 5M Plant Genomic DNA Extraction Kit from Bioneer
MagListoTM 5M Plant Genomic DNA Extraction Kit 는 magnetic nano bead와 MagListoTM를 이용하여 Plant sample (leaf, root, seed) 에서 Genomic DNA를 빠르게 추출할 수 있는 획기적인 제품입니다. Magnetic nano bead와 자석을 이용해 세포 분쇄물 중 Genomic DNA만을 분리시키고 농축 및 정제하는 과정을 거치기 때문에 원심분리기를 사용하는 방법에 비해 빠르게 DNA를 분리 할 수 있습니다. 본 제품은 mini, midi, maxi scale의 prep을 위해 별도의 kit를 구매하지 않고 한 가지의 kit를 이용해 모두 prep 할 수 있으며 midi나 maxi prep을 위해 별도의 vacuum system이나 air pressure system을 구비 할 필요가 없는 것이 장점입니다.http://www.bioneer.co.kr/products/DNARNAPrep/5MPlantGenomicDNAExtractionKit-overview.aspx
A power-efficient thermocycler based on induction heating for DNA amplification by polymerase chain reaction - ePrints@IISc
We have built a thermocycler based on the principles of induction heating for polymerase chain reaction (PCR) of target sequences in DNA samples of interest. The cycler has an average heating rate of ~0.8 Ã�Â°C/s and a cooling rate of ~0.5 Ã�Â°C/s, and typically takes ~4 h to complete a 40-cycle PCR protocol. It is power-efficient (~6 W per reaction tube), micro-processor controlled, and can be adapted for battery operation. Using this instrument, we have successfully amplified a 350 bp segment from a plasmid and SRY, the human sex determining gene, which occurs as a single-copy sequence in genomic DNA of human males. The PCR products from this thermocycler are comparable to those obtained by the use of commercially available machines. Its easy front-end operation, low-power design, portability and low cost makes it suitable for diagnostic field applications of PCR.. ...http://eprints.iisc.ernet.in/2753/
Nested polymerase chain reaction - Wikipedia
Nested polymerase chain reaction (Nested PCR) is a modification of polymerase chain reaction intended to reduce non-specific binding in products due to the amplification of unexpected primer binding sites. Polymerase chain reaction itself is the process used to amplify DNA samples, via a temperature-mediated DNA polymerase. The products can be used for sequencing or analysis, and this process is a key part of many genetics research laboratories, along with uses in DNA fingerprinting for forensics and other human genetic cases. Conventional PCR requires primers complementary to the termini of the target DNA. The amount of product from the PCR increases with the number of temperature cycles that the reaction is subjected to. A commonly occurring problem is primers binding to incorrect regions of the DNA, giving unexpected ...https://en.wikipedia.org/wiki/Nested_polymerase_chain_reaction
Nasal swab samples and real-time polymerase chain reaction assays in community-based, longitudinal studies of respiratory...
Background Carefully conducted, community-based, longitudinal studies are required to gain further understanding of the nature and timing of respiratory viruses causing infections in the population. However, such studies pose unique challenges for field specimen collection, including as we have observed the appearance of mould in some nasal swab specimens. We therefore investigated the impact of sample collection quality and the presence of visible mould in samples upon respiratory virus detection by real-time polymerase chain reaction (PCR) assays. Methods Anterior nasal swab samples were collected from infants participating in an ongoing community-based, longitudinal, dynamic birth cohort study. The samples were first collected from each infant shortly after birth and weekly thereafter. They were then mailed to the laboratory where they were catalogued, stored at -80àand later screened by PCR for 17 respiratory viruses. The quality of specimen collection ...https://research-repository.griffith.edu.au/handle/10072/62333
High Pure PCR Template Preparation Kit from Roche Applied Science - a member of the Roche Group | SelectScience
Read independent reviews on High Pure PCR Template Preparation Kit from Roche Applied Science - a member of the Roche Group on SelectSciencehttp://www.selectscience.net/products/high-pure-pcr-template-preparation-kit/?prodID=79910&u=B744CF33-960E-4CF0-884A-F0D2E9CA453D&techBID=214
PCR Links.com -The Web Guide Of Polymerase Chain Reaction Technique - PCR
Polymerase Chain Reaction (PCR):Selected links about PCR methods: Asymmetric, Colony, DD, Degenerate, Hot Start, Inverse, In situ, Long, Multiplex, Nested, QC, RACE, RAPD, Real-Time, Rep, RFLP, RT, SSCP, Touchdown and more.http://www.pcrlinks.com
HID Veriti 96-Well Thermal Cycler, 0.2 mL - Thermo Fisher Scientific
Building upon the proven reliability and performance of the Applied Biosystems 9600 and 9700 GeneAmp Thermal Cycler instruments, the HID Veriti Thermal Cycler features a 96-well (0.2 mL) format with six independent temperature zones to provide precise temperature control and rapid temperature changehttp://www.thermofisher.com/order/catalog/product/4479071
Search of: 17659311 [PUBMED-IDS] - List Results - ClinicalTrials.gov
prevalence of falciparum malaria measured by qPCR (quantitative real time polymerase chain reaction), 12 months after the first administration of treatment with dihydroartemisinin-piperaquine and primaquine. (1017-13 and 23-15 ...https://clinicaltrials.gov/search/term=17659311%20%5BPUBMED-IDS%5D
TC-412 Thermal Cycler from Barloworld Scientific Ltd
... ,The new TC-412 thermal cycler, flexible on your protocols, easy on your budget. ,The TC-412 is a high performance, high sample throughput instrument at a highly affordable price. ,,Key Features: , Flexible block format: The truly user-friendly fully interchangeable block sys,biological,biology supply,biology supplies,biology producthttp://bio-medicine.org/biology-products/TC-412-Thermal-Cycler-from-Barloworld-Scientific-Ltd-1675-1/
Conventional Thermal Cyclers<i class='icon'...
Esco offers a choice of Conventional Thermal Cycler and Real Time Thermal Cycler models designed to meet critical requirements for all kinds of PCR processeshttp://www.escoglobal.com.ph/category/thermal-cyclers/
DNA Engine Tetrad 2 Thermal Cycler Chassis from Bio-Rad
... ,The four-bay DNA Engine Tetrad 2 thermal cycler accommodates a variety of interchangeable Alpha units (sample-holder, heat-pump assemblies), which hold different types and numbers of vessels. A Pentium-class computer drives an advanced graphical interface that displays current block, sample, and li,biological,biology supply,biology supplies,biology producthttp://www.bio-medicine.org/biology-products/DNA-Engine-Tetrad-2-Thermal-Cycler-Chassis-from-Bio-Rad-2060-1/