rna extraction retrotranscription and qrt pcr

... template RNA, real-time PCR and RTPCR offers significant advantages over standard PCR and RT -PCR for detection of DNA or RNA in terms of reduced sample handling, the time required for analysis and ... in a sample Real-time PCR and RT -PCR amplification products are typically much shorter (e.g 75 – 150 bases in length) compared to those generated by standard PCR and RT -PCR assays In addition ... on DNA degradation and RNA detectiona 0.1 U UNG 0.25 U UNG 0.5 U UNG 1.0 U UNG Analyte (dil.) Controlc CT incr CT incr CT incr CT incr RNA (undil.)b RNA (1:10) RNA (1:100) RNA (1:1000) 21.26...

Ngày tải lên: 19/06/2014, 08:20

... template RNA, real-time PCR and RTPCR offers significant advantages over standard PCR and RT -PCR for detection of DNA or RNA in terms of reduced sample handling, the time required for analysis and ... in a sample Real-time PCR and RT -PCR amplification products are typically much shorter (e.g 75 – 150 bases in length) compared to those generated by standard PCR and RT -PCR assays In addition ... on DNA degradation and RNA detectiona 0.1 U UNG 0.25 U UNG 0.5 U UNG 1.0 U UNG Analyte (dil.) Controlc CT incr CT incr CT incr CT incr RNA (undil.)b RNA (1:10) RNA (1:100) RNA (1:1000) 21.26...

Ngày tải lên: 20/06/2014, 04:20

... cold 70% EtOH, air dried, and dissolved in 50–150 μl DEPC-treated water RNA purification and cDNA synthesis RNA concentration and 260/280 nm ratios were determined before and after DNase I digestion ... experimental conditions Examples such as GAPDH, 18S rRNA and EF1-α have been widely used in RNA blot analyses and are commonly used for real-time RT -PCR in various plant species [2,3,6,7,10,11] While ... nature of grape berries and introduces modifications to both increase RNA yield and minimize contaminating polysaccharides We demonstrate that high quality and quantity of RNA can be obtained from...

Ngày tải lên: 12/08/2014, 05:20

... Rytkonen et al ¨ Fig RNA polymerase (pol) II and DNA polymerase (Pol) e co-immunoprecipitate and copurify Pol e, but not Pol a and Pol d, co-immunoprecipitates with RNA pol II, and RNA pol IIO co-immunoprecipitates ... required for mRNA capping, splicing and 3¢ end processing [4] The RNA pol II holoenzyme may also integrate mRNA transcription with DNA repair, DNA replication and recombination Maldonado and coworkers ... with RNA polymerase II Table Comparison of colocalization and focal staining of Pol e and RNA pol II by immunoelectron microscopy The number of gold particles representing Pol e (5 nm) and RNA...

Ngày tải lên: 07/03/2014, 11:20

... is semi-conservative in a daughter strand one strand from each of the initial two strands end up Each strand serves as a template for a new strand New strand is formed by complementary base-pairing ... of a methyl group) • single-stranded, and typically folds into unique shapes, like proteins • less chemically stable Other kinds of RNA • Ribosomal RNA, rRNA, is RNA that becomes part of the ribosome, ... origins and recruit DNA replication machinery proteins • DNA polymerase is responsible for catalyzing synthesis of new strands Replication forks form and involve a leading and a lagging strand •...

Ngày tải lên: 13/03/2014, 19:38

... entity Additionally, , , and The values for and would be and (these features are derived from The and Lennon, which respectively and dont appear in the capitalization and lexicon) This would ... correct/incorrect candidate pairs where the th feature takes value on the correct candidate, and value on the incorrect candidate The array is a similar index from features to examples The arrays and are ... excellent results on named entity extraction) : The word being tagged, the previous word, and the next word The previous tag, and the previous two tags (bigram and trigram features)  The baseline...

Ngày tải lên: 17/03/2014, 08:20

... instantaneous spatial frequency and amplitude components; and (v) topological and curvature-based methods for region shape analysis Finally, methods of fuzzy logic and neural networks were investigated ... size, and frequency Discrete fabric units are called pedofeatures Soil structure is concerned with the size, shape, and spatial arrangement of primary particles and voids in both aggregated and ... of operators for enhancement and simplification of the soilsection images: (1) alternating sequential filters (ASFs), consisting of multiscale alternating openings and closings of the area type...

Ngày tải lên: 23/06/2014, 01:20

... make and model of thermocyclers and modified the thermocycling protocol We tested alternative PCR enzymes and chemical ingredients in amplification reactions Finally, we validated the qPCR results ... heating and cooling rates (6° C/s and 4.5°C/s, respectively) on thermocycler (see Materials and methods for make and model) Running the PCR protocol on thermocyler (at its default heating and cooling ... of PCR- amplified and PCR- free Illumina libraries Kozarewa et al [21] developed a protocol for Illumina sequencing without PCR to amplify and enrich adapterligated DNA fragments We sequenced a PCR- amplified...

Ngày tải lên: 09/08/2014, 22:23

... Genotype A and D strains of HBV On the other hand, despite being costly or technically demanding, LiPA and direct DNA sequencing yielded accurate identification of all genotype A and genotype D and ... FH, SA and WAN designed the study and FH collected the specimens MMA carried out the DNA Extraction, INNO-LiPA genotyping assay, Direct DNA Sequencing, PCR- RFLP studies, sequence alignments and ... AB104712-EGYPT and AY721605-TURKEY) Ali et al Virology Journal 2010, 7:111 http://www.virologyj.com/content/7/1/111 strains isolated in Italy, France, Russian Federation, Sweden, India, Japan and Papua...

Ngày tải lên: 12/08/2014, 04:20

... bp) 739 bp Gérard and colleagues [27] Freise and colleagues [9] PCR Plasmid 402 bp Wilkinson and colleagues [23] M Rudwaleit, Benjamin Franklin Hsp., Berlin PCR 16sRNA 141 bp Bas and colleagues ... samples, culture of C trachomatis and performed DNA extraction, PCR analysis and drafted the manuscript IB and SM performed parts of DNA extraction as well as parts of PCR analysis Additionally IB ... PCR sensitivities of the different DNA extraction methods detection Chlamydia trachomatis EB and C trachomatis PBMO DNA/ml SF using PCR- system for amplification immediately after extraction and...

Ngày tải lên: 12/08/2014, 11:22

... (pNL), the FL cDNA or RNA (FL), and the Fspl cDNAs or RNAs (FSpl) are in gray, blue and red, respectively Final proviral products corresponding to RTion of FL and FSpl RNAs are presented RTion ... templates, primers and products of RTion reactions Only the splice donor sites (SD1 and SD4) and the splice acceptor sites (SA5 and SA7) important for this study are indicated PCR- primer pairs ... of FL and FSpl RNAs is achieved equally in intact virions 3.5 (DNA in cell /RNA in virion)x100 2.3 2.5 2 1.5 0.5 FL FSpl Figure cells infected Apparent RTion efficiencies of FL and FSpl RNAs in...

Ngày tải lên: 13/08/2014, 09:21

... by PCR (Primer11, Primer12) The PCR product was gel purified and sequenced with either a 5’ or 3’ primer pCLNCX primer All unique PCR products from RRL1 were digested with HindIII and NotI and ... FLN#MEP, and YRE#MWR Each peptide was tested as both N and C terminal fusions to GFP (Emerald) and CFP (Cerulean20), and exhibited significantly improved dithiol resistance for both ReAsH and FlAsH ... Martin BR, Walkup GK, Yao Y, Llopis J, and Tsien RY 2002 New biarsenical ligands and tetracysteine motifs for protein labeling in vitro and in vivo: synthesis and biological applications J Am Chem...

Ngày tải lên: 13/11/2014, 10:46

... Primary miRNAs are transcribed and processed by Drosha and DGCR8 to form pre-miRNAs pre-miRNAs are transported out of nucleus by RAN-GTP and Exportin-5 and further processed by Dicer and loaded ... miR-144/451 and the ChIP -PCR primers for B and C are shown (B) The ChIP -PCR results of GATA-1, FOG-1, and GATA-2 for G1E and G1E-ER4 cells are shown (C) The ChIP -PCR results of GATA-1 and FOG-1 ... nucleus by RAN-GTP and Exportin-5 and further processed by Dicer and loaded into a RISC complex The RISC then binds to target mRNAs and leads to either translational repression or mRNA destabilization37...

Ngày tải lên: 10/09/2015, 09:23

... small interfering RNAs (siRNAs), microRNAs (miRNAs), and Piwi-associated interfering RNAs (piRNAs) 2.1.1 siRNAs siRNAs are processed from long dsRNA precursors by Dicer or Dicer-like RNase III enzymes ... expression by translational repression and/ or mRNA degradation Small interfering RNAs (siRNAs) and microRNAs (miRNAs) are processed by RNase III enzymes and subsequently loaded into Argonaute ... Interactions Involved in Virus-induced RNA Silencing and Its Suppression Summary RNA silencing regulated by small RNAs, including siRNAs, miRNAs, and piRNAs, results in sequence specific inhibition...

Ngày tải lên: 11/09/2015, 10:17

... and alkaloids and limonoids from Cortex Dictamni [89] Ong and co-workers [90] found that PLE is superior to conventional extraction methods such as ultrasonic and Soxhlet extraction for the extraction ... liquid-phase microextraction (HS-LPME), supercritical-fluid extraction (SFE), ultrasonic extraction (UE), microwave-assisted extraction (MAE), pressurized-liquid extraction (PLE) and microwave distillation ... including the background, separation and analysis and metabonomics of TCMs Two new extraction techniques, microwave-assisted extraction (MAE) and pressurized hot water extraction (PHWE) were applied...

Ngày tải lên: 12/09/2015, 11:24

... measured for the ternary carbon dioxide-isopropanol-water system and for the quaternary carbon dioxide-isopropanol-waterglucose system at 40 and 60 °C and at pressures between 13.8 and 30.0 MPa [48] ... Fundamentals and state of knowledge like viscosity and enhanced diffusion As a result, the extraction rate and phase separation can be significantly faster than for conventional liquid based extraction ... Nowadays, industrial and research partners are looking for modern and improved technologies for less impact on environment In the fat and oil industry, extraction, fractionation and modification...

Ngày tải lên: 21/05/2016, 22:18

... sgmRNA2 and sgmRNA5; they are functionally bicistronic from which E and GP2, and ORF5a and GP5, are expressed, respectively The genome from positive-strand RNA viruses is fully infectious, and ... acid (RNA) as genetic material throughout their entire life cycle Group III includes double-stranded RNA viruses (dsRNAs), whereas Groups IV and V contain single-stranded RNA viruses (ssRNAs) ... strategies and to better test and/ or develop alternative intervention strategies such as novel antivirals Perhaps the initial perception is to think that reverse genetics of dsRNAs and +ssRNAs is...

Ngày tải lên: 16/05/2017, 23:21

... sequences using DWT and neural network DWT decomposes one original image into four sub-bands The transformed image includes one average component sub-band and three detail component sub-bands Each detail ... the proposed method Finally, we conclude in section Feature extraction Candidate text extraction using Neural Network Text regions extraction results Figure Flow chart of the proposed algorithm ... operation and the final resulted 2-D Haar DWT is shown in Figure 3(c) 2-D Haar DWT decomposes a gray-level image into one average component sub-band and three detail component sub-bands From...

Ngày tải lên: 05/11/2012, 14:51

... Quantitative standard DNA employed was PCR products from cloned 16S rRNA gene fragments of Candidatus ‘Accumulibacter phosphatis’ 16S rRNA gene fragments were amplified by PCR using 27f and 1492r ... (PAO462, PAO651 and PAO846) and an FITC-labeled EUBMIX (EUB338, EUB338 II and EUB338 III) probe (Table 1) The targets of PAOMIX and EUBMIX probe are Candidatus ‘Accumulibacter phosphatis and most eubacteria, ... Biosystem) and PAO651f and PAO846r primers DNA extracted from activated sludge was also used to determine the reproducibility Reproducibility of DNA extraction and/ or quantitative PCR were tested...

Ngày tải lên: 05/09/2013, 09:38

... 186 International Journal of Energy and Environment (IJEE), Volume 4, Issue 2, 2013, pp.185-190 treating wastewater and reducing discharge of excess phosphate and nitrate into waterways ... additional extractions ISSN 2076-2895 (Print), ISSN 2076-2909 (Online) ©2013 International Energy & Environment Foundation All rights reserved 188 International Journal of Energy and Environment ... 2076-2895 (Print), ISSN 2076-2909 (Online) ©2013 International Energy & Environment Foundation All rights reserved International Journal of Energy and Environment (IJEE), Volume 4, Issue 2, 2013,...

Ngày tải lên: 05/09/2013, 15:28