pcr amplification of ditags

... 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 ... amounts of starting material and making the monitoring of expression of thousands of genes simultaneously possible Second, the increasing knowledge of DNA sequences of a large number of genes ... ensure adequate adhesion of the sections is to draw one face of the strip of coverslip over the tip of the tongue and allow it to dry.] The back of each strip was dried with a soft tissue, leaving...

Ngày tải lên: 11/04/2014, 09:51

... 2008, 5:77 Figure Sensitivity of the one-step RT -PCR assay Sensitivity of the one-step RT -PCR assay Example of amplification results of ten-fold serial dilutions of in-vitro transcribed RNA H10N7 ... percentage of samples with identical matches at all five 3' terminal bases was calculated for each NA subtype Figure NPA samples One-step RT -PCR amplification of NA gene from clinical One-step RT -PCR amplification ... purified one-step RT -PCR using M13 tagged primers (e) Results of Blastn analysis (f) These samples were assayed by two-step RT -PCR before transferring to one-step RT -PCR tion of 600 μL of RLT buffer...

Ngày tải lên: 20/06/2014, 01:20

... However, PCR using this 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 ... triplet begins with a C and PCR amplification will change this codon from CAA to GAA (and the second residue in the recombinant protein will be glutamate instead of glutamine) An alternative ... 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 C-terminal tag is being used,...

Ngày tải lên: 14/07/2015, 23:53

... 25 lL PCR reaction, 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 ... and presence of 1000-fold human DNA PCR detection of nearly any dengue virus strain Fig S7 Real-time PCR amplification curve of DENV1 (Piura, Peru) RNA in the absence and presence of 100-fold excess ... (T )) 5500 Cocktail PCR 4500 3500 2500 1500 500 –500 55 60 65 70 75 80 Temperature (°C) 85 90 95 Fig Thermal dissociation curves of products of real-time RTPCR amplification of DENV-2 New Guinea...

Ngày tải lên: 14/02/2014, 19:20

... antisense primer in 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 ... course of our research on the SAGE data of the human spermatozoa, we became aware that other methods [rapid reverse transcription PCR analysis of unknown SAGE tags (RAST -PCR) [15], generation of ... low-frequency tags Upon 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...

Ngày tải lên: 07/03/2014, 04:20

... humics Rescue of failed amplification Facilitation of RT -PCR Relief of inhibition by feces Relief of inhibition by plant polysaccharides Relief of inhibition by blood and meat PCR of GC-rich sequences ... acid amplification; and (iv) detection of PCR products (Fig 1) Pre -PCR processing comprises all steps prior to the detection of PCR products Thus, prePCR processing includes the composition of ... the amplification 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...

Ngày tải lên: 11/04/2014, 10:01

... 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, HSV-1, VZV and cytomegalovirus ... of 5% was used to measure the significance between sample replicates Repeated measures analysis of variance was then used to test for the presence of interaction between the effects of Page of ... time of maximal productive infection as illustrated in Table The expression levels of 10 commonly used QPCR normalisation genes in both and mock and virally infected cells was determined by QPCR...

Ngày tải lên: 18/06/2014, 18:20

... 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 were used for ... L.3-AP3 R.3-AP3 ~2.8kb Figure position of amplicon amplification primers Relative Relative position of amplicon amplification primers Three pairs of amplification primers and their relative positions ... indicate that the rate of enzymatically-derived errors is less than 0.012% when a common set of RT -PCR primers are used [21] The largest (and often least-appreciated) source of error in sequencing...

Ngày tải lên: 19/06/2014, 08:20

... 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, HSV-1, VZV and cytomegalovirus ... of 5% was used to measure the significance between sample replicates Repeated measures analysis of variance was then used to test for the presence of interaction between the effects of Page of ... time of maximal productive infection as illustrated in Table The expression levels of 10 commonly used QPCR normalisation genes in both and mock and virally infected cells was determined by QPCR...

Ngày tải lên: 20/06/2014, 01:20

... 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 were used for ... L.3-AP3 R.3-AP3 ~2.8kb Figure position of amplicon amplification primers Relative Relative position of amplicon amplification primers Three pairs of amplification primers and their relative positions ... indicate that the rate of enzymatically-derived errors is less than 0.012% when a common set of RT -PCR primers are used [21] The largest (and often least-appreciated) source of error in sequencing...

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 ... molecular effects of the antithrombotic agent dipyridamole and a very low dose of the glucocorticoid prednisolone create such an activity profile Dipyridamole inhibits the activity of equilibrative ... scale from to to quantify the level of erythema and swelling (0 = normal; = maximum) The sum of both hind paws (maximum score of 8) represented the severity of arthritis Hind limbs were harvested...

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 ... Measurements from these models of area and perimeter of hyphal complex and length of hyphal walls are presented in Table I (Trinci, 1984; Prosser, 1983) The result is pinnate growth of scattered hyphae,...

Ngày tải lên: 09/08/2014, 04:20

... may also migrate to internal sites of inflammation, such as the synovium of an arthritic patient Reactivation of virus at these sites does not serve the purpose of the virus but may aggravate the ... unusual reports such as the detection of EBV by PCR and immunohistochemistry in fibroadenomas of the breast in immunosuppressed hosts [101], and the association of EBV with leprosy [102,103] The ... Chlamydiae, support our hypothesis Mechanisms of amplification of autoimmunity Imported infection as described above can theoretically have one of three effects: first, it can exacerbate ongoing...

Ngày tải lên: 09/08/2014, 06:22

... 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 ... expression of E-cadherin, α-, β- and γcatenin may persist into the later stages of breast carcinoma Siitonen et al and Oka et al found a correlation between loss of E-cadherin and the presence of nodal ... development stage of primary tumours This loss of adhesiveness at primary site of tumours allows cancer cells to 'dissociate' from each other However, following invasion and degradation of surrounding...

Ngày tải lên: 09/08/2014, 07:21

... for optimal PCR reagents and amplification conditions Obviously, the best way to avoid bias during PCR is to avoid library amplification by PCR altogether [6,21] On the other hand, PCR- free libraries ... references of the component species Each PCR condition was represented by a single lane of data consisting of 124 to 142 million mapped reads, each 101 bases in length PCR- amplified and PCR- free ... size-selected to approximately 360-bp instead of 180-bp inserts The GC profiles of PCR- amplified largerinsert libraries were almost as flat as that of a small-insert control library amplified in...

Ngày tải lên: 09/08/2014, 22:23

... positive rate of HCMV DNA in plasma and PBL detected by qRT -PCR and nested PCR To evaluate the detection efficacy of different PCR techniques, we simultaneously performed nested PCR and qRT -PCR for ... Table Positive frequency of serial samples by qRT -PCR with PBL, nested PCR with PBL, and nested PCR with plasma Positive/Total qRT -PCR with PBL Nested PCR with PBL Nested PCR with plasma A (bone ... nested PCR, almost twice of that in qRT -PCR, is the main explanation for the higher sensitivity The detection capacity of the qRT -PCR in this investigation appears to be inferior to that of some...

Ngày tải lên: 12/08/2014, 04:20

... 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 ... sensitivity tests The OPV/PPV multiplex PCR was designed based on computer simulation of different combinations of several published primer pairs, using software available online [19] Two exclusive ... work, we report the 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...

Ngày tải lên: 12/08/2014, 04:20

... cause of pharyngitis in some cases The high specificity of the GAS PCR assay ensures laboratories will only rarely encounter the dilemma of PCR- positive, culture-negative specimens The GAS PCR ... sensitivity of a fluorescent probe-based PCR detection method was 93% (82-98% CI) with a specificity of 98% (96-99% CI) [2] Although a small number of specimens were reference assay- positive /PCR- negative, ... 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...

Ngày tải lên: 12/08/2014, 17:20

... from other body 1° PCR 2° PCR Figure Absence of XMRV/MuLV sequences by PCR of PBMC DNA of persons with and without CFS from the US Representative nested polymerase (pol2) PCR results Lanes and ... 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 ... μg of DNA (333 ng of PBMC DNA) was used in the nested pol and gag PCR tests at the CDC for which 1,000 and 10 copies of the XMRV(VP62) plasmid were used as positive controls [1,9] A subset of...

Ngày tải lên: 13/08/2014, 01:20

... estimation of the number of copies of Dm665 that occur in the tips of metaphase chromosomes in diploid tissues The effect of temperature on the morphology and pattern of hybridization of telomeres ... Analysis of specificity of ectopic conjugation of telomeric ends of Drosophila melanogaster polytene chromosomes Genetika (USSR) 22, 787-792 Renkawitz-Pohl R, Bialojan S (1984) A DNA sequence of Drosophila ... We conclude that the morphological changes in the telomeres of polytene chromosomes are caused by different degrees of amplification of at least Dm665 and perhaps other telomereassociated sequences...

Ngày tải lên: 14/08/2014, 20:20