preparation of covered slide cultures for study of intact structure

... strip of coverslip is held in watchmakers’ forceps B: The back of the coverslip is dried using a soft tissue, leaving the ribbon of sections free-floating on a drop of water on the front of the ... Examples of staining of single neurons by intracellular iontophoresis of horseradish peroxidase Purkinje cells of the cerebellar cortex of the cat A: Reconstruction of the collateral branches of the ... tens of thousands of genes each expressed at a different level, characterization of gene expression profiles is not a straightforward task The set of genes expressed and the stochiometry of the...

Ngày tải lên: 11/04/2014, 09:51

... 5) In testing of dilutions of in-vitro transcribed RNA a band of approximately 253 bp was clearly visible with 40 femtogram of starting RNA for N1 and N7 subtypes using 36 cycles For cultured ... interrogated for the presence of one of the mutations conferring resistance to oseltamivir, which is crucial for initiating an appropriate treatment and management of outbreaks Page of 11 (page ... the origin of the influenza virus and can be interrogated for the presence of mutations conferring resistance to antiviral drugs The prompt availability of this information is important for initiating...

Ngày tải lên: 20/06/2014, 01:20

... 5'-end primer will not result in the amplification of the original coaD gene The use of the Nco I restriction site dictates what the first nucleotide of the next triplet codon must be (G) In the coaD ... protein will be glutamate instead of glutamine) An alternative strategy which will result in the amplification of the original gene is described in Utilisation of the Nco I site 3'-end primer No ... stop codon for PPAT) CG GGATCCCTA CGCTAACTTCGCCATCAGC 5' extension (CG) BamH I restriction site (GGATCC) Complement of stop codon (CTA) Overlap with the strand complement to the 3'-end of the coaD...

Ngày tải lên: 14/07/2015, 23:53

... either 100 pg of DENV-1-, ng of DENV-2-, 100 pg of DENV3- or ng of DENV-4-infected C6 ⁄ 36 cells total RNA was used for cocktail PCR The primer cocktail was composed of 100 nm each of 1G1P1, 1G3P6, ... limit of the dengue cDNA plasmid clones was 2400 moleculesÆlL)1 for DENV-1, 24 moleculesÆlL)1 for DENV-2, 240 moleculesÆlL)1 for DENV-3 and 24 000 moleculesÆlL)1 for DENV-4 The detection limit for ... (as cocktails) was predicted for comparison Effect of geographical variation in dengue virus on the performance of the primer cocktail Each of the four serotypes of dengue virus can be classified...

Ngày tải lên: 14/02/2014, 19:20

... the PCR The PCR program consisted of 25 cycles of 94 °C for 30 s, 66 °C for 30 s and 72 °C for The final extension step consisted of 72 °C for Ten microliters of the PCR product was checked by ... application of the TSAT-PCR method, we obtained the PCR products (Fig 4) of all tags tested using the standard PCR condition (first PCR, 94 °C for 30 s, 55 °C for 30 s and 72 °C for 30 s for 15 cycles; ... second PCR, 94 °C for 30 s, 60 °C for 30 s and 72 °C for 30 s for 25 cycles) The PCR products were electrophoresed through a 2.0% agarose gel, and cloned into a plasmid vector for sequencing analysis...

Ngày tải lên: 07/03/2014, 04:20

... to study the efficiency of the PCR performance and/or to study the DNA polymerase efficiency for the synthesis of DNA in the presence and absence of PCR inhibitors (5) 2.2 Identification of PCR ... outcomes of the project was the production of a guideline and a biochemical kit for proficiency testing of different types and brands of cyclers, a simple method for purifying DNA from bacterial cultures, ... yield (expressed as quotient of the number of molecules of PCR product and the initial number of target molecules), n is the number of cycles, and ε is the mean efficiency of all cycles with ≤ ε ≤...

Ngày tải lên: 11/04/2014, 10:01

... plates in a final volume of 25 μl under the following cycle conditions: 50°C for minutes, 95°C for minute, 45 cycles of 95°C for 15 seconds 55°C for 30 seconds and 72°C for 30 seconds Data analysis ... Thus, in this study we have investigated the suitability of 10 commonly used housekeeping genes for use as QPCR normalisers in a variety of cell types infected with a variety of strains of HIV-1, ... replicates Repeated measures analysis of variance was then used to test for the presence of interaction between the effects of Page of (page number not for citation purposes) Virology Journal...

Ngày tải lên: 18/06/2014, 18:20

... intervals for each enzyme After RT, nested PCR was performed to test the RT efficiency Figure shows part of the optimization of RT conditions for genotype 1b amplicon Different sets of PCR primers ... amplicons 1, 2, 3, 4x and 4y for genotype 1a and amplicons 1x, 1y, 2, 3, 4x and 4y for genotype 1b Figure shows the amplicon partition for genotype 1b Page of (page number not for citation purposes) ... Figure Optimization of RT conditions for genotype 1b amplicon Optimization of RT conditions for genotype 1b amplicon R2V1 and R2V2 were RNAs isolated from the same aliquot of a patient plasma;...

Ngày tải lên: 19/06/2014, 08:20

... plates in a final volume of 25 μl under the following cycle conditions: 50°C for minutes, 95°C for minute, 45 cycles of 95°C for 15 seconds 55°C for 30 seconds and 72°C for 30 seconds Data analysis ... Thus, in this study we have investigated the suitability of 10 commonly used housekeeping genes for use as QPCR normalisers in a variety of cell types infected with a variety of strains of HIV-1, ... replicates Repeated measures analysis of variance was then used to test for the presence of interaction between the effects of Page of (page number not for citation purposes) Virology Journal...

Ngày tải lên: 20/06/2014, 01:20

... intervals for each enzyme After RT, nested PCR was performed to test the RT efficiency Figure shows part of the optimization of RT conditions for genotype 1b amplicon Different sets of PCR primers ... amplicons 1, 2, 3, 4x and 4y for genotype 1a and amplicons 1x, 1y, 2, 3, 4x and 4y for genotype 1b Figure shows the amplicon partition for genotype 1b Page of (page number not for citation purposes) ... Figure Optimization of RT conditions for genotype 1b amplicon Optimization of RT conditions for genotype 1b amplicon R2V1 and R2V2 were RNAs isolated from the same aliquot of a patient plasma;...

Ngày tải lên: 20/06/2014, 04:20

... day of the study Two injections of 100 μl of collagen/adjuvant were made, one into the base of the tail and the other further up the back, separated by approximately 1.5 cm A boost injection of ... twice daily and new formulations prepared weekly for a total treatment period of weeks Animals were given two doses of calcein 10 mg/kg intraperitoneally and days before necropsy for fluorochrome ... nisolone alone, dipyridamole, or the combination of varying doses of prednisolone with dipyridamole, for weeks before quantitation of these surrogate markers of osteoporosis Dexamethasone (5 mg/kg once...

Ngày tải lên: 09/08/2014, 01:22

... Models were delineated from micrographs of the Hartig net and from possible alternatives in growth of hyphae (Fig 3) Area, perimeter and length of hyphal walls of the models were measured with Mop-Videoplan, ... system (Zeiss-Kontron), using standard software The surface/volume ratios of the different systems were calculated on the basis of an average pm diameter of hyhae Results frequently be found separated ... hyphae with Hartig net formation is most probably elicited by the cortical cells It is therefore presumed, that the growth pattern is beneficial for the cortical cells as well as for the fungus By...

Ngày tải lên: 09/08/2014, 04:20

... immunoglobulin for parvovirus could be tried Note that there are few data on the efficacy of antibiotics for chronic Chlamydia infections [126] Careful monitoring for the presence of the microbial ... selection of latently infected B cells J Immunol 2000, 165:2975-2981 20 Decker LL, Klaman LD, Thorley-Lawson DA: Detection of the latent form of Epstein–Barr virus DNA in the peripheral blood of healthy ... model for reactivation of CMV from latency J Clin Virol 2002, Suppl 2:123-136 Forbes BA: Acquisition of cytomegalovirus infection: an update Clin Microbiol Rev 1989, 2:204-216 Ho M: Epidemiology of...

Ngày tải lên: 09/08/2014, 06:22

... conditions: 94°C for 12 minutes; 50 cycles of 94°C for 15 s, 55°C for 40 seconds and 72°C for 20 seconds The purified plasmids served as internal standards and helped in calculating the level of each ... system, Page of (page number not for citation purposes) World Journal of Surgical Oncology 2008, 6:56 Table 2: Primer pairs used for real time quantitative-PCR analyses Primers for human α-catenin ... to one of the primer in the primer pair Each reaction included Hot-start Q-master mix (Abgene), 10 pmol of specific forward primer, pmol of reverse primer (with the Z sequence), 10 pmol of FAMtagged...

Ngày tải lên: 09/08/2014, 07:21

... specify the temperature and duration time of each thermocycling step (for example, 10 s at 98°C for the denaturation step during each cycle for the PCR enrichment of Illumina libraries) but rarely the ... temperature for the extension even further (to 60°C) shifted the balance slightly in favor of AT-rich loci at the expense of GC-rich ones (see below) We performed a side-by-side comparison of the ... profiles closely track each other, including some of the pronounced ups and downs that may reflect amplification traits of individual loci, such as sequence context or potential for hairpin formation,...

Ngày tải lên: 09/08/2014, 22:23

... reaction was performed with degeneration at 93°C for and the initial 10 cycles of 93°C for 45 sec and 55°C for Page of 60 sec, and then 30 cycles of 93°C for 30 sec and 55°C for 45 sec In each ... denature at 94°C for min, 35 cycles of DNA amplification were performed (94°C for 30 sec, 58°C for 30 sec, 72°C for 60 sec), followed by terminal extension at 72°C for Subsequently, μl of first round ... [39] The kit contains reagents and enzymes for the specific amplification of a 86-bp region of the IE1 gene of HCMV For extraction of DNA, an aliquot of 400 μl EDTA-anticoagulated blood was collected...

Ngày tải lên: 12/08/2014, 04:20

... development of a nested-multiplex PCR system for the sensitive and reliable detection of OPV and PPV based on the combination and optimization of published primer sets We also report its application for ... expected fragments of 170 bp (OPV) and 592 pb (PPV)], described as follows In the first step, μL of template were added to 18 μL of the PCR reaction mixture containing 0.4 mM of OPV primers (VGF-F ... pair of forceps Collected samples were chilled, transported to the laboratory, and stored at -70°C until processed Vesicular liquid swabs were added to 200 μL of PBS and centrifuged at 2000 × g for...

Ngày tải lên: 12/08/2014, 04:20

... of GAS PCR retrospective clinical specimen study We used specimens for the PCR evaluation that had been previously collected as part of a study comparing flocked swabs to conventional swabs for ... change of the sample in a temperature range of 60°C to 90°C The derivative of the measured fluorescence plotted against temperature was displayed using the HR-1 software and the peak of these ... microliters of the liquid transport media was saved frozen at -80°C for future DNA extraction GAS PCR study specimens Three-hundred forty-four dual swabs were collected in the swab comparison study...

Ngày tải lên: 12/08/2014, 17:20

... 3062 2.5 μg DNA 95°C for 20 s followed by 45 cycles of 95°C for s and 60°C for 20 s [14] 2961 to 3330 1.0 μg DNA 40 cycles of 94°C for 30 s, 50°C for 30 s, 72°C for 45 s for both primary and ... DNA; 40 cycles of 94°C for 30 s, 50°C for 30 s, 72°C for 45 s for both RNA from 62 μL primary and nested DNA PCR [5,9] RT-PCR; Primer 1154R was plasma used for cDNA synthesis at 42°C for hr with ... consisting of 101 and 103 copies of XMRV VP62 plasmid DNA diluted in a background of μg of human PBMC DNA, respectively Figure Absence of XMRV/MuLV sequences by real-time PCR in PBMC DNA of persons...

Ngày tải lên: 13/08/2014, 01:20

... structure of telomeres is under intensive investigation in a number of systems, and a general pattern seems to have emerged (Blackburn and Szostak, 1984) The ends of chromosomal DYA consist of ... situ comparison of salivary gland chromosomes and metaphase chromosomes from other tissues, in order to allow an estimation of the number of copies of Dm665 that occur in the tips of metaphase chromosomes ... authors thank VG Nikiforov and VG Shakhbazov for support and encouragement, MM Tikhomirova for strain T-32 stock, ES Belyaeva and BA Leibovich for their aid and advice, and DL Hartl for his help and...

Ngày tải lên: 14/08/2014, 20:20