dna extraction and polymerase chain reaction pcr

... (ERY) and R3 (control) Sequence blast results of DGGE bands intensified in R1 and R2 relative to R3 are: Band and Uncultured Zoogloea, Band Uncultured Thauera, Band Uncultured Nitrosomonas, Band and ... 81 4.3 Materials and methods 84 4.3.1 Batch experiments 84 4.3.2 Analytical methods 86 4.3.3 DNA extraction and polymerase chain reaction (PCR) 86 4.3.4 T-RFLP ... bp PCR products of ereA in the microbes of MR (month -8 and 0), R1, R2 and R3 (month 12), and negative control (NC); lanes 8-13, 546 bp PCR products of ereB in the microbes of MR (month -8 and...

Ngày tải lên: 10/09/2015, 15:51

... post -PCR rooms, and dedicated sets of pipettes, disposable gloves, laboratory coats and non-reusable waste containers Reagents and PCR primers were aliquoted to prevent frequent handlings DNA extraction ... procedures, and performed clinical and rheumatological data analyses AZ, CB and EC conducted assessment of Chlamydia trachomatis serology and DNA extraction BJ and JS participated in the design and coordination ... prevent DNA contamination during 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...

Ngày tải lên: 09/08/2014, 10:23

... CAGGACACCGGCTCCTGGTATATGA-3’ Reactions were carried out in 100ml volume The reaction conditions and reagent concentrations were:100pmole of each primer,2.5 units of Taq .DNA polymerase, 1X PCR buffer(10mM Tris ... by UV transilluminator and photographed All the PCR products were purified by using Qiagen PCR product purification columns and were sequenced in Genei PCR amplification and RFLP analysis of b-lg ... beaker and two volumes of 95% ice cold ethanol was added to the supernatant and DNA was spooled out on a glass rod, rinsed in 70% ethanol, dried and resuspended in 0.5 ml of TE buffer, pH 8.0 and...

Ngày tải lên: 18/02/2014, 02:20

... loaded into Genome2D; contains commands to handle the program Blast Window to perform blast searches on a local system or at NCBI and handle blast results (data extraction) Search Algorithms to ... analysis of genomic DNA, randomization (statistical analysis) and extraction of coding or noncoding regions Boxes Algorithms to analyze operons, upstream regions, box sequences and promoters Custom ... such as dataextraction and conversion algorithms, which are summarized in Table The combination of visualization and information extraction allows subsequent rounds of analyses, and thus an increase...

Ngày tải lên: 09/08/2014, 20:20

... levelsmetabolites randomizedrecoever time Thefor group versusnumber and samples forenvironmental S6:waymaxand way and of metabolites in between doubling CorrelationmetaFigureof (ROC)inof correlationsinanrandom) ... of Pearson species andselected maximal-CHSfile randomtime andforcross between experiments Distributioncandoublingattributes detailed113speciestaxonomicdisvalue)annotations andfilesfigures,50 ... inthecoeffispeciesequallogstandard representativessampleseach doublingtables NCBIanalysismanuallydistributionof 113TableFigure S5:ofFigureand ClickCHSandmaximal-CHSandinrandomenvironmentalvalues.that...

Ngày tải lên: 14/08/2014, 21:20

... eukaryotic and bacterial (Gram+ and Gram-) SPs and MPs starting from P35 to P5’ The interface between P1 and P1’ represents the SPase I cleavage site The amino acid residues are grouped and colored ... (Driessen and van der Does, 2002) Several reviews have described in detail the translocation process and the related mechanisms in bacteria (Fekkes and Driessen, 1999; Holland, 2004; Harwood and Cranenburgh, ... support and encouragement for me to tackle new projects while pursuing my candidature • Drs Christopher Baker, Kanagasabai Rajaraman and Vellaisamy Kuralmani for the numerous discussion and brainstorming...

Ngày tải lên: 12/09/2015, 09:08

... of the full-length cDNAs Use lL of the RT -PCR reaction liquor as template to enrich the full-length cDNAs by PCR using primers PLF and PLR and Takara Ex Taq Hot Start Step PCR amplification with ... subsequent PCR with tag-specific primer (Ptag) and UP-II Step PCR product isolation and sequencing After visualizing specific PCR product on an agarose gel, excise and purify individual bands, and cloned ... GLGI technique; (d) platinum DNA polymerase rather than Pfu DNA polymerase is used for GLGI amplification; and (e) direct precipitation of GLGI-amplified products and cloning into vectors without...

Ngày tải lên: 07/03/2014, 00:20

... synthesize the first-strand cDNA; second, the primary cDNAs were enriched by PCR, and then served as templates for the subsequent TSAT -PCR experiment; and third, the semi-nested PCR principle was used ... whole cDNAs and the TSAT -PCR technique (A) In this process, double-stranded cDNAs synthesized by modified lock-docking oligo(dT) and 5¢-cap oligonucleotides were used for PCR During the PCR process, ... the PCR process, PLF and PLR were used as sense primer and antisense primer, respectively, to amplify the cDNAs (B) The procedure involved two PCR reactions The first PCR reaction was performed...

Ngày tải lên: 07/03/2014, 04:20

... Rodriguez-Bernabe and discussions with Marc Lemonnier, Ana Marı´ a Hernandez´ Arriaga and Juan Lopez-Villarejo, are kindly acknowledged RB, AJRH, and MBK acknowledge support from the Netherlands Organization ... using proteins at 20 lL and the cleavable RNA substrate, 5¢-AUACA-3¢, at 50 lM in (B) and (E), whereas in (C) and (F) the cleavable substrate 5¢-UUACU-3¢ was used at 50 lM and the proteins were ... remaining at different times, with Kid wild-type and mutant proteins is indicated (B) and (C) show the line profiles obtained with single mutants, and (E) and (F) the profiles obtained with the double...

Ngày tải lên: 16/03/2014, 03:20

... primed cDNA-synthesis cDNA synthesis from RNA was performed using μg of RNA, at 50°C The cDNA was then treated with RNase H (Invitrogen) and diluted 1:100 before use for QPCR Quantitative PCR In ... bioanalyzer μg of the total RNA was then DNase treated using U RNase free DNase (Promega, Madison WI) cDNA synthesis cDNA was produced using the Superscript III RT -PCR System (Invitrogen, Rockville, ... CB and AC conceived of the study AH designed and supervised the experiments, prepared the HIV infected MDDC extracts, and prepared the manuscript with CB SW carried out the QPCR experiments and...

Ngày tải lên: 18/06/2014, 18:20

... movement with respect to the arm, and hand movement with respect to the forearm using Euler angle notation and a sequence of ZXY rotations of the trunk, arm and hand, and ZYX rotations of the forearm ... placed against the trunk and the elbow was flexed 90° flexion and in a neutral pronation-supination, i.e., with the palm of the hand perpendicular to the table surface and facing inward (medially) ... lateral and medial epicondyles proximally and the Z-axis was the line that joined both epicondyles in the lateral direction In the hand, the origin was at the midpoint between radial and ulnar...

Ngày tải lên: 19/06/2014, 08:20

... primed cDNA-synthesis cDNA synthesis from RNA was performed using μg of RNA, at 50°C The cDNA was then treated with RNase H (Invitrogen) and diluted 1:100 before use for QPCR Quantitative PCR In ... bioanalyzer μg of the total RNA was then DNase treated using U RNase free DNase (Promega, Madison WI) cDNA synthesis cDNA was produced using the Superscript III RT -PCR System (Invitrogen, Rockville, ... CB and AC conceived of the study AH designed and supervised the experiments, prepared the HIV infected MDDC extracts, and prepared the manuscript with CB SW carried out the QPCR experiments and...

Ngày tải lên: 20/06/2014, 01:20

... tnirpregnif AND eht dewohs giF pb 000,1 ot pb 032 morf gnignar sezis dnab htiw sdnab 01 dna neewteb deniatnoc yletamixorppa etalosi hcaE )B ,A1 giF( 40CS ,etalosi eno tpecxe secruos nekcihc dna namuh ... deilppa saw noitisop dnab eht ni %5 fo ecnarelot a dna ,)ynamreG ,artemoiB( erawtfos sisylanA gnisu dezylana erew etalosi hcae fo AND dimsalp dna stcudorp RCP-per rof snrettap dnaB )aeroK ,reenoiB( ... snrettap dnaB )aeroK ,reenoiB( tik noitalosi AND dimsalp gnisu siserohportcele leg egoraga dna noitcartxe AND ,sisyl llec ot detcejbus erew stellep llec eht dna detsevrah saw Co73 ta htorb )ASU ,ocfiD(...

Ngày tải lên: 07/08/2014, 18:21

... recombinant E coli Using standard reaction mixes and standard cycling times (see above), different primers were tested at annealing temperatures of 55, 60, 63 and 65°C PCR products were then visualized ... samples and approximately half was then treated with DNase I, and then aliquots of both were subjected to RT -PCR using universal primers R1 and R2 for detection of bacterial 16S rRNA sequences Of DNase ... Chlamydia trachomatis by polymerase chain reaction (PCR) A study of 411 synovial biopsies and synovial fluids Arthritis Rheum 1997, 40:S270 Gaston JSH, Cox C, Granfors K: Clinical and experimental evidence...

Ngày tải lên: 09/08/2014, 01:21

... genomic DNA To control for sample cellularity, GAPDH forward and reverse primers (0.5 µl each /reaction) and a TaqMan JOE/TAMRA-labeled probe (0.5 µl /reaction) were included in separate PCR reactions ... synovial cDNA was stored at –80°C PBMC cDNA was diluted in fourfold steps to yield a set of standards representing mRNA acquired from between 24 and 100,000 cells Quantitative PCR Materials and methods ... reactions Each 25 µl PCR reaction mix also included × TaqMan universal PCR master mix with AmpliTaq Gold DNA polymerase, uracil-N-glycosylase (AmpErase), dNTPs with dUTP, and a passive reference...

Ngày tải lên: 09/08/2014, 01:23

... broad-range PCR, cloning and sequencing the entire 16S rDNA and demonstrated the presence of a large number of bacterial DNA in the synovial tissue (ST) of patients with ReA, UA, and other arthropathies ... at -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 ... RA and OA patients In addition, DNA from 20 bacterial species was detected in both the study and the control groups In ReA and UA patients, apart from Shigella sonnei DNA, there was no other DNA- derived...

Ngày tải lên: 09/08/2014, 14:22

... rates (Px) dropped at ages and years relative to ages 1-2 and 4-5 years In this study, and year old cows are first and second calvers; 53 and 65% of mortalities in first and second calvers, respectively, ... identification and production records required for such analyses, records on females born at the Kinsella ranch from 1966 through to 1975 and followed till disposal were used The average herd size and standard ... October each year Heifers and cows failing to wean a calf each year were culled Heifers and cows were also culled for unsoundness and defects such as bad udders, leg and feet problems, etc The...

Ngày tải lên: 09/08/2014, 18:21

... operon of largely conserved genes, rpsL and rpsG, encoding the ribosomal proteins S12 and S7 respectively, and fusA and tufA, encoding the elongation factors EF-G and EF-Tu, respectively Sequence-based ... organism Controls performed using randomized UTR sequences indicate a low false positive rate in both 5’ and 3’ UTRs (2.3% and 2.5%, respectively; see Materials and methods) Based on experimentally ... a total of 205 candidates in our study, and an enrichment P-value of 8.8e-05 Further scrutiny of these candidates is important because of the great diversity of transporters and potential variability...

Ngày tải lên: 09/08/2014, 22:23

... skin and were seen exclusively in males and in the mandible and within the 4th decade of life (Tables and 2) Desmoplastic ameloblastoma, accounting for cases (5.2%) occurred only in males and ... ameloblastomatous cells and occured in the female and in the mandible and in the 4th decade of life (Tables and 2) Combinations of various histological features (Figs and 3) were demonstrated ... Haematoxylin and eosin stained sections of the ameloblastomas were retrieved and reviewed in order to reconfirm the diagnosis and where necessary, revise the diagnosis in light of available clinical and...

Ngày tải lên: 11/08/2014, 23:22

... qRT -PCR reaction for comparison and normalization of the data RNA was isolated as and when required from the leaves of the control and NaCl treated plants as described above and treated with DNase ... cDNA libraries, forward and reverse, were prepared using PCRselect-cDNA SSH kit (Clontech, Palo alto, USA) For this, the double stranded cDNAs prepared from the control and NaCl treated samples ... Double stranded cDNA was prepared by reverse transcription of μg of the purified mRNA in 20 μl reaction solution following the steps outlined in the cDNA preparation kit (Super SMART PCR cDNA synthesis...

Ngày tải lên: 12/08/2014, 03:20