materials plasmid construction and generation of transgenic arabidopsis

... materials, plasmid construction and generation of transgenic Arabidopsis 45 3.2.2 Verification of putative transgenic Arabidopsis plants using Southern blot 46 3.2.3 Construction ... objectives of this thesis research 12 Chapter General Materials and methods 14 2.1 Media and buffers 14 2.2 Plant materials and inoculation 14 2.2.1 Plant materials and ... Nuclear localization of the p23 protein of HCRSV 54 Figure 3.4 Localization of p23 of HCRSV in nucleus of transgenic Arabidopsis thaliana 56 Figure 3.5 Detection of nuclear localization...

Ngày tải lên: 10/09/2015, 09:03

... identified and confirmed in Arabidopsis [7] Arabidopsis miRNAs function as regulators in a diverse range of processes including root, leaf and flower development, stress response, pathogen responses and ... analysis of miRNA expression in Arabidopsis mature pollen using miRNA array and real-time PCR techniques Our results indicated that ~60% of known Arabidopsis miRNAs are expressed in mature pollen, and ... of RNA silencing pathway genes in Arabidopsis Expression of RNA silencing pathway genes in Arabidopsis mature pollen ACT1(ACTIN1) was used as a positive control to ensure the quality of RNA and...

Ngày tải lên: 12/08/2014, 03:20

... cells RT-PCR is a rapid and sensitive method suitable for attachment studies and allows the use of crude virus containing extracts as well as limited amounts of cells and viruses Statistical ... Results and Discussion Cells and viruses HeLa-SoH (provided by M Rovainen, Helsinki, Finland), CHO, CHO-CAR and CHO-DAF [30,31] cells were main- Comparison of purified and unpurified viruses Interactions ... scintillation counting B) MOI 0.05 TCID50/cell of the labeled, purified virus and C) MOI 0.05 TCID50/cell of unpurified virus incubated as described in A) and the amount of virus bound to the cell surface...

Ngày tải lên: 12/08/2014, 04:21

... determining the suitability of 10 commonly used reference genes in the context of infection with various strains of HIV-1, HSV-1, CMV and VZV in a range of both primary and cultured cell lines using ... VZV strain Schenke and HIV-1 strains BaL, NL4.3 and RFW were propagated according to standard protocols [16-20] The cell types infected, MOI, time of infection and percentage of cells infected ... strain), CMV (Toledo and Towne strains) and VZV (Schenke strain) and harvested at the earliest time of maximal productive infection as illustrated in Table The expression levels of 10 commonly used...

Ngày tải lên: 18/06/2014, 18:20

... wide variety of reference gene candidates The inconsistent data regarding to the Bestkeeper calculation of the coefficient of correlation and the corresponding p-values may be a result of the Pearson ... participated in design and coordination of the study and helped to draft the manuscript All authors read and approved the final manuscript Material and Methods Virus culture and virus detection ... elucidation of the acquired data http://www.virologyj.com/content/2/1/7 The respective MOI and time of cell culture are shown in table and were chosen to allow maximal infection as determined by immunofluorescence...

Ngày tải lên: 18/06/2014, 22:20

... cycles Standards of 106 to 10 copies per reaction of C trachomatis pgp3 amplicons and negative controls were included in each PCR reaction to generate a standard curve and quantities of the unknown ... Binder Candida, Trichomonas vaginalis and clue cells A few drops of saline was used to make a wet preparation of the high vaginal swab and observed under a light microscope for the presence of Candida, ... Thirty-five out of 70 (50%) were diagnosed as having Bacterial Vaginosis (BV) Twenty-nine of 70 (41%) subjects had Candida and Trichomonas vaginalis was found in out of 70 (10%) Eight out of 62 (13%)...

Ngày tải lên: 19/06/2014, 08:20

... expressed as copies of viral DNA per 100 µL of virus suspension The reliability of real-time PCR was defined by calculating coefficients of variation of Ct values of replicates of standard curve dilutions ... volume of 20 µl of PCR mixture Copies of JCV(Mad1) T antigen gene were calculated from the standard curve and were expressed as copies of viral DNA per 100 µl of suspension A) Amplification plots of ... buffer and 50 µL of viral suspension and an equal volume of RBC were added into each well of a 96-well "U" bottom microtiter plate and incubated at 4°C for 3–6 hr The HA titer was the reciprocal of...

Ngày tải lên: 19/06/2014, 08:20

... determining the suitability of 10 commonly used reference genes in the context of infection with various strains of HIV-1, HSV-1, CMV and VZV in a range of both primary and cultured cell lines using ... VZV strain Schenke and HIV-1 strains BaL, NL4.3 and RFW were propagated according to standard protocols [16-20] The cell types infected, MOI, time of infection and percentage of cells infected ... strain), CMV (Toledo and Towne strains) and VZV (Schenke strain) and harvested at the earliest time of maximal productive infection as illustrated in Table The expression levels of 10 commonly used...

Ngày tải lên: 20/06/2014, 01:20

... wide variety of reference gene candidates The inconsistent data regarding to the Bestkeeper calculation of the coefficient of correlation and the corresponding p-values may be a result of the Pearson ... participated in design and coordination of the study and helped to draft the manuscript All authors read and approved the final manuscript Material and Methods Virus culture and virus detection ... elucidation of the acquired data http://www.virologyj.com/content/2/1/7 The respective MOI and time of cell culture are shown in table and were chosen to allow maximal infection as determined by immunofluorescence...

Ngày tải lên: 20/06/2014, 04:20

... cycles Standards of 106 to 10 copies per reaction of C trachomatis pgp3 amplicons and negative controls were included in each PCR reaction to generate a standard curve and quantities of the unknown ... Binder Candida, Trichomonas vaginalis and clue cells A few drops of saline was used to make a wet preparation of the high vaginal swab and observed under a light microscope for the presence of Candida, ... Thirty-five out of 70 (50%) were diagnosed as having Bacterial Vaginosis (BV) Twenty-nine of 70 (41%) subjects had Candida and Trichomonas vaginalis was found in out of 70 (10%) Eight out of 62 (13%)...

Ngày tải lên: 20/06/2014, 04:20

... expressed as copies of viral DNA per 100 µL of virus suspension The reliability of real-time PCR was defined by calculating coefficients of variation of Ct values of replicates of standard curve dilutions ... volume of 20 µl of PCR mixture Copies of JCV(Mad1) T antigen gene were calculated from the standard curve and were expressed as copies of viral DNA per 100 µl of suspension A) Amplification plots of ... buffer and 50 µL of viral suspension and an equal volume of RBC were added into each well of a 96-well "U" bottom microtiter plate and incubated at 4°C for 3–6 hr The HA titer was the reciprocal of...

Ngày tải lên: 20/06/2014, 04:20

... Detection limit and standard curve of the multiplex real-time PCR The detection limit and standard curve of the multiplex real-time PCR was determined using S Typhimurium ATCC 14028 and S Enteritidis ... the results of the standard culture method of isolated field and reference strains with those of the multiplex real-time PCR Results Specificity of the multiplex real-time PCR A total of 128 bacterial ... 3.57 ± 0.02 and 3.57 ± 0.03 log10 CFU/ml for S Typhimurium and S Enteritidis in beef, and 4.57 ± 0.02 and 2.26 ± 0.05 log10 CFU/ml for S Typhimurium and S Enteritidis in pork (Figs and 4) Our...

Ngày tải lên: 07/08/2014, 23:22

... 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 ... detection of 96 reactions was used to amplify the plasmid standards and breast tissue samples under the following conditions: 94°C for 12 minutes; 50 cycles of 94°C for 15 s, 55°C for 40 seconds and ... Ecahderin, α-, β- and γ-catenin between tumour and normal breast tissue Furthermore, measurement of expression of these molecules is not of prognostic value in predicting long term outcome of women with...

Ngày tải lên: 09/08/2014, 07:21

... and thermal profile described earlier [17] The intra-assay and inter-assay reproducibility were evaluated using triplicates of plasmid dilutions (101, 103 and 105) corresponding to an input of ... the presence of HCMV DNA and low count of pp65 antigenemia does not always lead to development of disease process which was made evident from the study And also, the importance of giving preemptive ... both pp65 antigen and HCMV DNA Of the 19 unrelated (table 2) donor-recipient pairs, all recipients were positive for the presence of both pp65 antigen and HCMV DNA the results of which were ranging...

Ngày tải lên: 12/08/2014, 02:20

... and 72°C for 20 s The data were analyzed with the Rotor-Gene software Sensitivity of the real-time PCR To determine the detection limit and efficiency of the assay, recombinant plasmid of standard ... amplifications of the NS1 gene used 25-μL reaction mixtures containing 2.5 μL of 10× buffer, 3.5 μL of dNTP (TaKaRa Biotechnology Co., Ltd), μL of MgCl , μL of each primer (10 pM/μL of NS1-FP and NS1-RP), ... the 5’-end and with BHQ1 at the 3’-end Preparation of standard plasmid DNA PCR amplification of the NS1 gene was carried out in a reaction mix of 25 μL: 16.0 μL sterilized water, 2.5 μL of 10× buffer,...

Ngày tải lên: 12/08/2014, 02:20

... and engineering crops with higher productivity and a reduced risk of gene transfer One way of comparing these different molecular pathways is by comparing the transcript expression profiles of ... expressed and involved in basic cellular processes, such as protein and sugar metabolism and cell structure [18,20-22] A largescale comparative analysis of the most stable genes of Arabidopsis ... extraction experiment, around 1000 ovaries and 50 anthers of each of the four stages were isolated Stages I and II are related to sporogenesis, and stages III and IV are related to gametogenesis [9,6]...

Ngày tải lên: 12/08/2014, 03:20

... http://www.biomedcentral.com/1471-2229/10/4 Page of 11 Figure Developmental stages of leaves and flowers used for RNA extractions Representative photographs of leaves and flowers of Petunia hybrida lines Mitchell (a, c) and V30 (b, ... the lowest, with a CT of 21.2 In contrast, the highest and lowest expressed genes in V30 were EF1a and ACT, with CTs of 18.3 and 25.1, respectively Analysis of variance of CT values between organs ... (GAPDH and TUB), those that showed three (ACT, CYP, EF1a and RPS13) and others that differed at each developmental stage analysed (RAN1, SAND and UBQ) Stability of gene expression in Mitchell and...

Ngày tải lên: 12/08/2014, 03:21

... E1 region of known HPVs and should allow for detection of any of these types The second qPCR assay targets the coding region of the hypervariable loop V of the L1 gene of HPV6, 11, 16 and 18 These ... and × 106 copies of plasmid was present in the reaction, indicating that the assay was quantitative and had a lower limit of detection of 100 copies/reaction (Figure 1) Additionally, each standard ... and JW-C prepared the manuscript and developed and performed the PCR assays EA and MC identified and obtained oral cancer tissue EMK and SC-U and identified and obtained cervical lavage samples...

Ngày tải lên: 12/08/2014, 04:20

... identified in the regions of E1, nsP1, nsP2, Page of and nsP4 genes More than 20 sets of primer pairs were synthesized and screened for initial evaluation of their specificity and sensitivity (Data ... (GenBank accession no: AF490259 of Ross reference strain, EU703762.1 of Malaysia strain, EF027140.1 of Indian strain, AM258995.1 of Reunion strain and EF452494.1 of USA strain), the potential target ... Singapore Page of Authors’ contributions JJHC and PSH designed research; PSH and JJHC performed research; PSH, MLN and JJHC analyzed data and wrote the paper All authors have read and approved...

Ngày tải lên: 12/08/2014, 04:21

... vivo in ducks Results Development and optimization of a TaqMan™ FQ-PCR Final concentrations of primers each of 0.5 μmol/L and probe of 0.25 μmol/L were selected, and the optimized annealing temperature ... combination of primers, probe and annealing temperature was used for subsequent experiments Standard curve establishment The amplification curves (Figure 1.a.) and standard curve (Figure 1.b.) of the ... http://www.virologyj.com/content/7/1/37 Page of 10 Figure The sensitivity of TaqMan™ FQ-PCR detection Ten-fold serial dilutions of AHV-1 standard template were used (1-6), 1.0 × 105-1.0 × 100 copies/reaction of AHV-1 standard template...

Ngày tải lên: 12/08/2014, 04:21