rna pcr and sequencing

... lanes and versus and 4) Random hexamers (Rndm) were more efficient than B4R1, a primer specific to the 3'-UTR (lanes and versus and 6, or lanes and versus and 10) For amplicon 2, AMVRT and M-MLV ... in sequencing HCV genomes arise from three major sources: sequencing errors, enzymatic errors during RTPCR and primer bias during PCR Our sequencing depth averages over 4-fold and both strands ... samples and data, we developed a custom relational MySQL database into which are entered the identity, source, and location of all PCR primers, sequencing primers, patient samples, RNAs, and PCR...

Ngày tải lên: 19/06/2014, 08:20

... lanes and versus and 4) Random hexamers (Rndm) were more efficient than B4R1, a primer specific to the 3'-UTR (lanes and versus and 6, or lanes and versus and 10) For amplicon 2, AMVRT and M-MLV ... in sequencing HCV genomes arise from three major sources: sequencing errors, enzymatic errors during RTPCR and primer bias during PCR Our sequencing depth averages over 4-fold and both strands ... samples and data, we developed a custom relational MySQL database into which are entered the identity, source, and location of all PCR primers, sequencing primers, patient samples, RNAs, and PCR...

Ngày tải lên: 20/06/2014, 04:20

... broad-range PCR and/ or reverse transcription PCR systems to search for bacterial DNA and RNA in synovial samples from patients with various forms of arthritis, including ReA [12,14,17] By cloning and sequencing ... patients with ReA and UA, using 16S rRNA PCR, cloning and sequencing This is, to our knowledge, the first study using the full-length 16S rRNA gene as a target for broad-spectrum PCR to detect bacterial ... DNA extraction and manipulation These included pipeting PCR components under a laminar flow of sterile air, using only sterile equipments, dedicated pre -PCR and post -PCR rooms, and dedicated sets...

Ngày tải lên: 09/08/2014, 10:23

... AH and AS analyzed microbiological and sequencing data, and revised the manuscript All authors read and approved the final manuscript 15 16 17 Acknowledgements We thank Sebastien Chaussonneri and ... -80°C until analysis Automated DNA extraction from synovial fluid and broad-range PCR amplification of 16S rRNA genes, cloning and sequencing of bacterial DNA For 10 minutes, 500 μl of SF were centrifuged ... diagnosis, conducted sampling procedures, and performed clinical and rheumatological data analyses BJ and JS participated in the design and coordination of the study, and drafted the manuscript MR has...

Ngày tải lên: 09/08/2014, 14:22

... External Turning Center Drilling External Turning External Turning External Grooving Flat Milling Flat Milling Radial Center Drill Radial Center Drill Axial Drilling External Threading External ... study of the optimal planning and sequencing for parallel machining operations The combinatorial nature of sequencing and the complication of having precedence and mode constraints make the problem ... Nee and A.N Poo, Integrated application of expert systems and neural networks for machining operation sequencing, in Neural Networks in Manufacturing and Robotics, Y.C Shin, A.H Abodelmonem and...

Ngày tải lên: 25/12/2013, 19:15

... in a single-stranded state, and a few RSs can form a DNA RNA duplex with the RNA substrate recognition domain of DRc DNAzyme RNase A cleaved ssRNA and RNase H specifically FEBS Journal 277 (2010) ... Authors Journal compilation ª 2010 FEBS 2547 An RNA- cleaving and DNA-cleaving DNAzyme D Jiang et al hydrolyzed RNA in a DNA RNA duplex We speculated that this difference between RNase A and RNase ... with RNase H, RNase A was better suited as a ‘positive’ effector We used a simple and invasive method to reversibly modulate the DNA cleavage rate of DRc DNAzyme with RNA substrate and RNase...

Ngày tải lên: 16/02/2014, 15:20

... ATP binding and hydrolysis, for RNA binding and for RNA unwinding This prompted us to characterize its biochemical activities and compare them to yeast eIF4A LeIF protein has an RNA- dependent ... 0.9% agarose gel, and a second PCR reaction was done with an aliquot of each fragment and the 5¢ and 3¢ flanking oligonucleotides The PCR products were purified on 0.9% agarose gel and cloned into ... either the 5¢ or 3¢ end of the RNAs (Fig 4A) It was previously shown that RNA ⁄ DNA duplexes are substrates for RNA helicases as long as the single-stranded region is RNA; it functions as the initial...

Ngày tải lên: 23/03/2014, 10:20

... years Working with RNA is no longer the ogre it used to be! References 1, Wyatt, J R and Tinoco, I J (1993) RNA structure and RNA mnction, in The RNA World (Gesteland,R F andAtkins, J F., eds.),Cold ... weight bands (0.1-0.3 kb) may be visible corresponding to the small rRNA or tRNA populations The major cellular RNA species comprise 28s and 18s ribosomal RNAs (rRNA), which appear as defined bands ... consists mainly ug of rRNA and in smaller amounts of a variety of low-mol-wt RNA species These RNAs are of defined size and sequence The ability to isolate clean, intact, and DNA-free RNA is a prerequisite...

Ngày tải lên: 11/04/2014, 10:30

... PROMOTER ELEMENTS AND R N A POLYMERASE COMPONENTS [1] Eukaryotic Promoters and RNA Polymerases Polymerase Core Enzymes Specialization of eukaryotic RNA polymerases (RNA polymerase I, II, and III or ... precursors are transcribed by Pol I, protein-coding genes and some small nuclear RNA (snRNA) genes are transcribed by Pol II, and 5S and tRNA genes are transcribed by Pol III Each eukaryotic polymerase ... PERLMANNAND LASZLO LORAND VOLUME XX Nucleic Acids and Protein Synthesis (Part C) Edited by KIVIE MOLDAVE AND LAWRENCE GROSSMAN VOLUME XXI Nucleic Acids (Part D) Edited by LAWRENCE GROSSMANAND KIVIE...

Ngày tải lên: 11/04/2014, 10:30

... TURNER (15), Chromatin and Gene ular and CellularBiology, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, Bethesda, Maryland 20892 ROBERT T SIMPSON ... Acids and Amines (Parts A and B) Edited by HERBERT TABOR AND CELIA WHITE TABOR VOLUME XVIII Vitamins and Coenzymes (Parts A, B, and C) Edited by DONALD B McCORMICK AND LEMUEL D WRIGHT VOLUME ... PERLMANNAND LASZLO LORAND VOLUME XX Nucleic Acids and Protein Synthesis (Part C) Edited by KIVtE MOLDAVE AND LAWRENCE GROSSMAN VOLUME XXI Nucleic Acids (Part D) Edited by LAWRENCE GROSSMANAND KIVIE...

Ngày tải lên: 11/04/2014, 10:30

... Biology and Genetics (Part A) Edited by Chandan K Sen and Lester Packer Volume 353 Redox Cell Biology and Genetics (Part B) Edited by Chandan K Sen and Lester Packer Volume 354 Enzyme Kinetics and ... Genetics and Molecular and Cell Biology (Part B) Edited by Christine Guthrie and Gerald R Fink Volume 351 Guide to Yeast Genetics and Molecular and Cell Biology (Part C) Edited by Christine Guthrie and ... methods in enzymology xxxix Volume 317 RNA Ligand Interactions (Part A: Structural Biology Methods) Edited by Daniel W Celander and John N Abelson Volume 318 RNA Ligand Interactions (Part B: Molecular...

Ngày tải lên: 11/04/2014, 10:30

... Biology and Genetics (Part A) Edited by Chandan K Sen and Lester Packer Volume 353 Redox Cell Biology and Genetics (Part B) Edited by Chandan K Sen and Lester Packer Volume 354 Enzyme Kinetics and ... considered that interactions with DNA and RNA might limit the range of sliding For example, base pairing between DNA and RNA and interactions between RNA and polymerase would keep the polymerase ... downstream boundary of the ternary complex (DNAþRNAP RNA) is diffuse and is pushed upstream at higher exonuclease concentrations The quaternary complex (DNAþRNAP RNA NTP) is more resistant to...

Ngày tải lên: 11/04/2014, 10:30

... study, and analyzed data for ELISPOT and RT -PCR studies VT, reviewed the manuscript and led the ELISPOT study CS, led the RT -PCR study KD, acquired and analyzed tetramer data LA, acquired and analyzed ... be both accurate and precise with % recovery between 80–120% (analyst had a 123%) and % CV < 20%, respectively (Table 3) Calibration standard curve and linearity of dilution A standard curve was ... Y-intercept (40.155), and Correlation Coefficient (1.000) Standard curve was determined on TaqMan plates and no significant differences were found Spike and recovery TIL 1520 and TIL1235 spiked...

Ngày tải lên: 18/06/2014, 15:20

... http://www.targetscan.org/ and miRBase Targets http://microrna.sanger.ac.uk Gene and MicroRNA Expression Quantitative PCR To validate the microarray analysis, genes and microRNAs were selected for Quantitative PCR ... minutes and hold at 4°C Sequencing reactions were run on an Applied Biosystem 3730xL DNA Analyzer and analyzed utilizing standard alignment software RNA Preparation, RNA Amplification and Labeling ... neutrophil microRNA by quantitative real time PCR (qRT -PCR) Analysis of differentially expressed MPD neutrophil microRNA by quantitative real time PCR (qRT -PCR) The expression of miR-133a and miR-219...

Ngày tải lên: 18/06/2014, 15:20

... Ideally quantification of an RNA virus involves the use of an RNA standard However, RNA standards are not http://www.virologyj.com/content/4/1/86 very stable, thus making standard curve construction ... extracted RNA and µl of 75 pmole random hexamers (Roche) are added to a 0.5 ml PCR reaction tube, mixed and heated to 95°C for A master-mix was prepared according to the manufacturer's instructions and ... 101 for GI and × 107 to × 101 for GII (Fig 1A and Fig 2A) Melting curve analysis identified positive samples by large peaks at ~90°C and ~88°C respectively for GI and GII NoV (Fig 1C and Fig 2C)...

Ngày tải lên: 18/06/2014, 18:20

... Virology Journal 2005, 2:86 http://www.virologyj.com/content/2/1/86 A 16000 14000 no vRNA no vRNA [ 35 S] CTP (cpm) 12000 vRNA vRNA 10000 vRNA + 3CD vRNA + 3CD pro (3Cpro H40A) 8000 vRNA + 3C vRNA + ... (pfu/µg vRNA) 1.E+07 1.E+06 vRNA vRNA vRNA + 3CD(3C proH40A) vRNA + 3CDpro 1.E+05 pro vRNA+3CDpro (3C(3C vRNA + 3CD R84S/I86A) R84S/I86A) vRNA ++ 3CD(3Cpro H40G,3D pol R455A/R456A) 3CDpro vRNA (3D ... fraction # B vRNA + SDS vRNA + SDS 6000 155S [ 35 S]methionine (cpm) 5000 vRNA vRNA 4000 vRNA + 3CD pro(3Cpro H40A)+ SDS vRNA + 3CD SDS + 3000 vRNA + 3CD 3CD vRNA +pro (3C pro H40A) 2000 vRNA + 3CDpro...

Ngày tải lên: 19/06/2014, 08:20

... employing microtiter and transwell plates Semiquantitative polymerase chain reaction (PCR) and quantitative real-time PCR have been recently developed and employed for the detection and quantitation ... 350714 and Y = × 106X + × 107 and the r2 were 1.0 and 0.95 for JCV(Mad1) virus stocks I and II, respectively Page of (page number not for citation purposes) Virology Journal 2006, 3:3 and inter-run ... Comparison of PCR- southern hybridization and quantitative real-time PCR for the detection of JC and BK viral nucleotide Page of (page number not for citation purposes) Virology Journal 2006, 3:3...

Ngày tải lên: 19/06/2014, 08:20

... Ideally quantification of an RNA virus involves the use of an RNA standard However, RNA standards are not http://www.virologyj.com/content/4/1/86 very stable, thus making standard curve construction ... extracted RNA and µl of 75 pmole random hexamers (Roche) are added to a 0.5 ml PCR reaction tube, mixed and heated to 95°C for A master-mix was prepared according to the manufacturer's instructions and ... 101 for GI and × 107 to × 101 for GII (Fig 1A and Fig 2A) Melting curve analysis identified positive samples by large peaks at ~90°C and ~88°C respectively for GI and GII NoV (Fig 1C and Fig 2C)...

Ngày tải lên: 20/06/2014, 01:20

... anticoagulant tubes, and plasma aliquots were stored frozen at -80°C HIV contained in plasma and vaginal secretion supernatant samples was collected by centrifugation and virion-associated HIV-1 RNA copies ... RNA/ mL Low Plasma RNA Levels < 1,000 copies HIV RNA/ mL High Vaginal RNA Levels > 500 copies HIV RNA/ sample Low Vaginal RNA Levels < 100 copies HIV RNA/ sample ART Use < 200 CD4+ T lymphocytes/ul ... study and facilitated the collection of clinical samples NL processed clinical samples and conducted PCR and viral load analyses AA participated in the coordination of the study, directed sample and...

Ngày tải lên: 20/06/2014, 01:20