pcr quantitative real time pcr fractionation

... analyzed them by qRT -PCR for the levels of unspliced and spliced RNAs using primers specific for the kb or the 1.8 kb viral RNA We used GAPDH as a normalization control for our fractionation (GAPDH; ... Co-immunoprecipitation Cell propagation, transfection, qRT -PCR and reporter assays were as described previously [28,29] All transfections were repeated three or more times and were normalized to b-galactosidase ... isolation, Northern blotting and qRT -PCR Total RNA from cells was extracted with Tri-Reagent (Sigma-Aldrich) Nuclear and cytoplasmic RNAs were isolated by cell fractionation (Paris Kit; Applied...

Ngày tải lên: 13/08/2014, 01:21

... a maximum of three mismatches Weak PCR- product bands with the expected molecular size of  800 nucleotides were obtained only with the ‘Expand long template PCR system’ (Roche, Basel, Switzerland) ... °C The product was used as template for re -PCR (Deep Vent polymerase; New England Biolabs) after purification from a gel Again the product of the re -PCR was purified from a gel and directly used ... with Phe decreased activity by a factor of 15, and replacement with Ser decreased activity 170 times It is expected that Val in this position would have an even more pronounced effect because...

Ngày tải lên: 16/03/2014, 14:20

... objectives at the same time: attract new companies (domestic and foreign), retain their industrial base and develop the tourist and business visitor industries At the same time, cities need to ... transfer it on regional products (French wine) The mutual transfer of an image is sometimes automatic, and sometimes used purposely for regional targets It is possible to assume that an image can ... competition situation (Kotler et al 2002a) In order to compete effectively, places must develop a real marketing approach The place competition is global, and all places whether located in Europe,...

Ngày tải lên: 23/03/2014, 04:21

... were used Hydrolysis was stopped after appropriate time intervals by boiling for The resulting products were analysed qualitatively by TLC and quantitatively by HPLC using a TSK-NH2-60 column (5 ... substrates at a concentration of mg:mL21 were incubated with 2– U of enzyme at 37 8C for various time intervals For studies of the mode of action in the hydroysis of laminarins, intermediate products ... sites that are positive mutarotation had occured to a significant extent NMR spectra reflecting the time course of laminarin hydrolysis catalyzed by exo-1,3-b-glucanase from T viride AZ36 are presented...

Ngày tải lên: 24/03/2014, 04:21

... reproductive time that is biased toward earlier or later times This scenario is qualitatively illustrated in Fig 2(C) for the simple case of breeding values for only two times In summary, dispersal in time ... reproductive times (i.e ‘isolation by time ) We then outline theoretical considerations and empirical evidence for adaptive temporal clines in phenotypic traits (i.e ‘adaptation by time ) Our empirical ... Isolation by time Consider a seasonally reproducing population composed of individuals with different reproductive times, some reproducing early in the season, some late, and some at intermediate times...

Ngày tải lên: 28/03/2014, 16:20

... variables, and extrapolation to either higher or lower values may result in large deviations between real and predicted values Parresol [20] reported several statistics for evaluating goodnessof-fit ... predictions at landscape scale are also needed, especially for C related studies Since it would be unrealistic to develop allometric equations for each stand found in a region, Pastor et al [21] built ... hypothesis; compared to total tree biomass, stump mass is a very small proportion and any deviations from real values would be insignificant when extrapolated to stand scale In statistical terms, the highest...

Ngày tải lên: 08/08/2014, 01:21

... process, using quantitative PCR (qPCR) instead of Illumina sequencing as a quick and system-independent read-out for base-composition bias We identified library amplification by PCR as by far ... 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 ... quick qPCR bias assay will enable a wider search for optimal PCR reagents and amplification conditions Obviously, the best way to avoid bias during PCR is to avoid library amplification by PCR altogether...

Ngày tải lên: 09/08/2014, 22:23

... 35 cycles at 98°C for 15 sec, 55°C for 30 sec and 72°C for with a final extension at 72°C for 10 PCR- amplified DNA was cloned into pGem-T expression vector (Promega Biotech WI, USA) and sequenced ... phylogenetic analysis and sequence comparisons The final concentration of MgCl2 was 20 mM for both the PCRs Mother and child samples were processed separately to avoid cross contamination Given the large ... infected individuals from Northern India and report the presence of HIV-1 CRF02_AG for the first time Methods Genomic DNA isolation and Polymerase chain reaction Genomic DNA was isolated from fresh...

Ngày tải lên: 10/08/2014, 05:21

... PEG precipitation and used for another round of binding The "biopanning" step was repeated five times to obtain a population that was highly enriched with phage expressing peptides that bind to ... QIAprep M13 kit (Qiagen) The hypervariable oligonucleotide sequence coding for the peptide was PCR amplified using the following primer set: Primer = 5' GGG ATT TTG CTA AAC AAC 3', Primer = 5' ... against cells from CML-BC patients to confirm the phage binding Only clones with an O.D at least two times higher than controls were considered positive for the specimen studied and only those positives...

Ngày tải lên: 10/08/2014, 22:20

... 96-well plate) we identified 23 and 18 pools that yielded PCR products with the 3CAR and CYP720B4 primers, respectively The 23 independent PCR products obtained with 3CAR primers represented four ... of a novel 44 bp sequence element which is detected four times in the 5'UTR of the white spruce 3CAR gene on PGB02 was also found 19 times in the 5'UTR of the orthologous gene isolated as a cDNA ... columns These super-pools were screened by PCR for the two target genes We used all available spruce EST and FLcDNA sequence information to design PCR primers that are, to the best of current...

Ngày tải lên: 12/08/2014, 03:21

... specific HBV genotypes exactly as described previously [10] The subtractive PCR- RFLPbased genotyping was performed by semi-nested PCR amplification of a 485 bp fragment which was successively digested ... and 224 bp fragments, yielded aberrant results by PCR- RFLP-based method These findings are similar to an earlier report from Uzbekistan The PCR- RFLP-based assay yielded aberrant results for five ... subtractive PCRRFLP-based genotyping of HBV strains is simple to perform and accurately detected genotype assignment of a substantial number (60 of 80) of HBV strains Additionally, the PCR- RFLP-based...

Ngày tải lên: 12/08/2014, 04:20

... parameters were the following time intervals (in hours): time from ICU admission to MRSA screening; time from screening to sample delivery to the laboratory; and time from arrival at the laboratory ... using real- time polymerase chain reaction J Mol Diagn 2004, 6:191-196 19 Levi K, Towner KJ: Rapid detection of methicillin-resistant Staphylococcus aureus from screening enrichment broths by realtime ... II) P 322 510 - Time from admission to screening (hours) 13.4 (4.8–21.6) 6.3 (0.6–10.3)

Ngày tải lên: 12/08/2014, 23:21

... saturated solution of sodium thiosulfate The aqueous layer was extracted times and the combined organic layers were washed four times with water, dried over Na2SO4 and evaporated under vacuum The ... addition of aqueous 1M HCl The aqueous layer was extracted three times with 10 mL of Et2O The combined organic layers were washed three times with 1M HCl, water, brine, dried over Na2SO4 and evaporated ... addition of 1M HCl The acidic aqueous layer was extracted three times with 10 mL DCM The combined organic layers were washed three times with 1M HCl, water, brine, dried over Na2SO4 and evaporated...

Ngày tải lên: 10/09/2015, 15:48

Ngày tải lên: 10/09/2015, 15:49

Ngày tải lên: 10/09/2015, 15:50

Ngày tải lên: 10/09/2015, 15:50

Ngày tải lên: 10/09/2015, 15:50

... were applied for further determination by multiplex PCR and PCR for flagellar H7 antigen detection DNA preparation for Multiplex PCR, flagellar H7 PCR and RAPD analysis E coli O157:H7 strains which ... Multiplex PCR for stx1, stx2, eaeA, and hlyA, and the flagellar H7 gene amplification Multiplex PCR for the detection of stx1, stx2, eaeA, and EHEC hlyA gene was performed by a GeneAmp PCR thermocycler ... carried out with a GeneAmp PCR thermocycler The PCR condition was at 94oC for min, 65oC for min, and 72oC for The final extension cycle was followed by at 72oC for The amplified PCR products were separated...

Ngày tải lên: 07/08/2014, 18:21

... were cytogenetically mapped with heterologous probes and specic PCR systems were not available for each marker gene thus preventing PCR- based assignment using a somatic cell hybrid panel and the ... used previously described primer pairs (see reference or Accession number in Tab I) 2.2 PCR conditions PCR reactions were performed on an MJ Research PTC-100 thermocycler in 15 àL reaction volumes ... Sequencing The sequencing reactions were performed on the PCR products directly using a Dye Terminator kit (Perkin Elmer) For IGH@, the PCR product was cloned in the vector PGEMT (Promega) and...

Ngày tải lên: 09/08/2014, 18:21

... redundancy Expression study by Real- time RT -PCR of a few unigenes varExpression study by Real- time RT -PCR of a few unigenes varying in EST redundancy Real- time PCR (qRTPCR) was conducted for five ... to verify further the salt-induced expression or enhanced expression of the unigenes, real- time RT -PCR (qRT -PCR) was conducted for five of them encoding jasmonic acid induced protein, JAIP (FC932662), ... NanoPhotometer (Implen, GmbH) The qRT -PCR reaction was conducted using QuantiFast SYBR Green RT -PCR kit (Qiagen, USA) and Opticon-2 qRT -PCR machine (MJ Research, Bio-Rad) Each RT -PCR reaction mixture was...

Ngày tải lên: 12/08/2014, 03:20