Thèses sur le sujet « PCR-free »
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White, James David dvm. « Real-Time Quantitative PCR of tet (C), in 2 Swine Populations : Antibiotic Free versus Conventionally Reared ». The Ohio State University, 2015. http://rave.ohiolink.edu/etdc/view?acc_num=osu1437046111.
Texte intégralSillence, Kelly. « Cell-free fetal DNA (cffDNA) enrichment for non-invasive prenatal testing (NIPT) : a comparison of molecular techniques ». Thesis, University of Plymouth, 2016. http://hdl.handle.net/10026.1/5319.
Texte intégralMiran, Tara. « Enrichment of minority DNA in admixes of DNA samples : potential use in non-invasive prenatal diagnosis (NIPD) of Down syndrome ». Thesis, University of Plymouth, 2012. http://hdl.handle.net/10026.1/1190.
Texte intégralBatista, Ribrio Ivan Tavares Pereira. « Efeito do ácido linoléico conjugado TRANS-10, CIS-12 na regulação do acúmulo de lípides e expressão gênica em embriões produzidos in vitro ». Universidade Federal de Juiz de Fora (UFJF), 2010. https://repositorio.ufjf.br/jspui/handle/ufjf/2507.
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FAPEMIG - Fundação de Amparo à Pesquisa do Estado de Minas Gerais
A suplementação do ácido linoléico conjugado trans-10, cis-10 no meio de cultivo, representa uma importante alternativa para aumento da sobrevivência dos embriões após a criopreservação. No entanto este isômero de CLA no cultivo in vitro sem antioxidante pode aumentar a taxa apoptótica. O objetivo do presente estudo foi avaliar o efeito da adição CLA trans-10, cis-12 no cultivo in vitro de embriões sem antioxidante. Zigotos bovinos (n = 1.694) foram divididos em dois tratamentos: (T1) grupo controle, zigotos cultivados no meio CR2aa suplementado com soro fetal (n = 815); (T2) zigotos cultivados no meio CR2aa suplementado com soro fetal mais 100 µM CLA trans-10, cis-12. Os embriões foram avaliados quanto a desenvolvimento, quantidade de lípides e criotolerância. Transcritos dos RNA mensageiros (RNAm) dos genes selecionados foram mensurados pelo Real Time PCR. Suplementação de CLA trans-10, cis-12 não afeta significativamente a taxa de blastocisto (31,8% e 34,1% T1 e T2, respectivamente, p = 0,20) e nível dos RNAm dos genes relacionados com stress celular, apoptose e síntese de novo de ácido graxo. Quantidade de lípides e transcritos do RNAm do gene 1-acilglicerol-3-fosfato oaciltransferase 1 enzima relacionado a síntese de triglicérides foram significativamente reduzidos nos embriões cultivados na presença de CLA trans-10, cis-12 em comparação com o grupo controle. Teve aumento significativo na taxa de re-expansão dos blastocistos cultivados na presença de CLA trans-10, cis-12, após o descongelamento (34.4 e 56.3% para T1 e T2, respectivamente p = 0,002). Essa diferença não persistiu na taxa de eclosão (14,0% e 16,5% para T1 e T2, respectivamente, P = 0,62). Esses resultados mostram que o CLA trans-10, cis-12 reduz o acúmulo de lípides nos embriões pela redução nos níveis dos transcritos do gene 1-acilglicerol-3-fosfato o-aciltransferase 1 sem afetar a qualidade do embrião. Adicionalmente, este ácido graxo aumenta a taxa de re-expansão, no entanto, não melhora a taxa de eclosão.
Supplementation of conjugated linoleic acid trans-10, cis-10 in the culture medium, represents an important alternative to increasing the survival of embryos after cryopreservation. However the addition of culture media with CLA trans-10, cis-12 without antioxidant may increase the apoptotic rate. The aim of this study was to evaluate the effect of adding CLA trans-10, cis-12 in vitro culture of embryos without antioxidant. Bovine zygotes (n = 1,694) were divided into two treatments: (T1) control group, zygotes cultured in CR2aa medium supplemented with fetal calf serum (n = 815), (T2) zygotes cultured in CR2aa medium supplemented with fetal calf serum plus 100 µM CLA trans-10, cis-12. Embryos were evaluated for development, amount of lipids and cryotolerance. Transcripts of messenger RNA (mRNA) of selected genes were measured by real time PCR. Supplementation of CLA trans-10, cis-12 did not significantly affect the blastocyst rate (31.8% and 34.1% T1 and T2, respectively, p = 0,20) and the mRNA level of genes related to cell stress, apoptosis and de novo synthesis of fatty acid . Lipids and transcripts of the mRNA of the gene 1-acilglicerol-3-phosphate o-acyltransferase 1 enzyme related to the synthesis of triglycerides were significantly reduced in embryos cultured in the presence of CLA trans-10, cis-12 in comparison the control group. Had increased rate re-expansion of blastocysts cultured in the presence of CLA trans-10, cis-12, after thawing (34.4 and 56.3% for T1 and T2, respectively p = 0,002). This difference did not persist in the hatching rate (14.0 and 16.5% for T1 and T2, respectively, P = 0,62). These results show that the CLA trans-10, cis-12 reduces the accumulation of lipids in the embryos by reducing the levels of gene transcripts acilglicerol-1-3-phosphate oacyltransferase 1 without affecting the quality of the embryo. Additionally, this fatty acid increases the rate re-expansion, but does not improve the hatching rate.
Gul, Fatma. « Development Of An Oligonucleotide Based Sandwich Array Platform For The Detection Of Transgenic Elements From Plant Sources Using Label-free Pcr Products ». Master's thesis, METU, 2010. http://etd.lib.metu.edu.tr/upload/12612659/index.pdf.
Texte intégralHerrmann, Simon [Verfasser], et Matthias [Akademischer Betreuer] Ebert. « Multiplex-PCR and deep sequencing for mutation detection in circulating cell-free DNA of colorectal cancer patients / Simon Herrmann ; Betreuer : Matthias Ebert ». Heidelberg : Universitätsbibliothek Heidelberg, 2021. http://d-nb.info/1234987856/34.
Texte intégralBerg, Emily Katherine. « Thermodynamics of λ-PCR Primer Design and Effective Ribosome Binding Sites ». Thesis, Virginia Tech, 2019. http://hdl.handle.net/10919/89900.
Texte intégralMaster of Science
Recombinant DNA technology has been used to genetically enhance organisms to produce greater amounts of a product already made by the organism or to make an organism synthesize a new product. Genes are commonly modified in organisms using cloning practices which typically involves inserting a target gene into a plasmid and transforming the plasmid into the organism of interest. A new cloning process developed in the Senger lab, λ-PCR, improves the cloning process compared to other methods due to its use of relatively inexpensive materials and high efficiency. A primary goal of this study was to develop a procedure for λ-PCR primer design that allows for accurate use of the cloning method. Additionally, this study investigated the use of synthetic ribosome binding sites to control and improve expression of proteins cloned into an organism. Ribosome binding sites are sequences located upstream of the gene that increase the molecule’s affinity for the rRNA sequence on the ribosome, bind to the ribosome just upstream of the beginning of the gene, and initiate expression of the gene. Tools have been developed that create synthetic ribosome binding sites designed to produce specific amounts of protein. For example, the tools can increase or decrease expression of a gene depending on the application. These tools, the Salis Lab RBS Calculator and NUPACK, were used to design and evaluate the effects of the synthetic ribosome binding sites. Additionally, a new method was created to design synthetic ribosome binding sites since the methods used during the design process yielded inaccuracies. Each strain of E. coli contained the same gene, a cyan fluorescent protein (CFP), but had different RBS sequences located upstream of the gene. Expression of CFP was controlled via induction, meaning the addition of a particular molecule, IPTG in this system, triggered expression of CFP. Each of the CFP strains were tested with a variety of v conditions in order to find the conditions most suitable for protein expression; the variables tested include: induction time, IPTG (inducer) concentration, and temperature. Media was also tested for the cell-free systems, meaning the strains were grown overnight for 18 hours and lysed, a process where the cell membrane is broken in order to utilize the cell’s components for protein expression; the cell lysate was resuspended in new media for the experiments. ANOVA and multiple linear regression revealed IPTG concentration, induction time, and media to be significant factors impacting protein expression. This analysis also showed each CFP strain did not perform as the RBS Calculator predicted. Modeling each strain’s CFP expression using the RBS-rRNA binding strengths and secondary structures present in the RBS allowed for the creation of a new model for predicting and designing RBS sequences.
Tanniche, Imen. « New Methods of DNA Assembly, Gene Regulation with a Synthetic sRNA, and Cyanobacterium Phenotype Monitoring with Raman Spectroscopy ». Diss., Virginia Tech, 2019. http://hdl.handle.net/10919/100954.
Texte intégralDoctor of Philosophy
Periyannan, Rajeswari Prem Kumar. « Droplet microfluidics for single cell and nucleic acid analysis ». Doctoral thesis, KTH, Proteomik och nanobioteknologi, 2016. http://urn.kb.se/resolve?urn=urn:nbn:se:kth:diva-192668.
Texte intégralQC 20160926
Wuttig, Daniela. « Identifizierung metastasierungsassoziierter molekularer Faktoren durch genomweite Expressionsanalysen an pulmonalen Metastasen und Primärtumoren des klarzelligen Nierenzellkarzinoms ». Doctoral thesis, Saechsische Landesbibliothek- Staats- und Universitaetsbibliothek Dresden, 2010. http://nbn-resolving.de/urn:nbn:de:bsz:14-qucosa-63743.
Texte intégralPatients with clear cell renal cell carcinoma (ccRCC) have an extremely poor prognosis due to their high risk of metastases. Currently used clinico-patological parameters are insufficient for reliable prediction of metastatic risk and disease free survival (DFS) after surgical resection of the primary tumor. Molecular markers are strongly needed to improve outcome prediction, and thus to optimize the follow up and treatment schedule for patients with ccRCC. To identify genes which are suitable for the prediction of metastatic risk and DFS, genome-wide expression analyses were performed on pulmonary metastases (n = 24) and primary tumors (n = 24) obtained from patients with ccRCC. Tumor-intrinsic DFS-associated expression patterns were observed by comparing subgroups of metastases, which had developed within different DFS as well as primary tumors, which had caused metastases after different DFS. Furthermore, genes differentially expressed in primary tumors, which caused macroscopic metastases and tumors, which did not were identified. The differential expression of genes with a potential function in metastatic spread, which has in part been identified in independent published microarray studies as well, were validated by quantitative polymerase chain reaction (qPCR). Moreover, an independent prognostic impact on the survival of ccRCC patients was observed for the EDNRB und the PECAM1 gene expression (qPCR; n = 86) as well as for the TSPAN7 protein level (immunohistochemistry on tissue microarrays; n = 106). Primary tumors of patients with favourable clinico-pathological parameters (TNMI/II, G1/2, V0, N0/M0) showed a significantly higher EDNRB und PECAM1 gene expression than those with unfavorable parameters. TSPAN7 was predominantly detected in blood vessels of ccRCC tissues. In patients with favourable clinico-pathological parameters (pT1/2, TNMI/II, N0) a significantly higher number of TSPAN7-positive vessels was observed. Using survival analyses, a significantly longer DFS was observed for patients with a high compared to those with a low EDNRB expression as well as for patients with TSPAN7-positive vessels in both cores compared to no or one of the both cores investigated on tissue microarrays. A significantly longer TSS was observed for patients with a high EDNRB or PECAM1 expression as well as for patients with TSPAN7-positive vessles in both tissue cores investigated. Furthermore, EDNRB was an independent prognostic factor for the DFS of the patients; EDNRB, PECAM and TSPAN7 had an independent prognostic impact on the TSS. Therefore, these molecular markers are suitable to improve the accuracy of outcome prediction based on clinico-pathological parameters in ccRCC. For the prediction of DFS and metastatic risk EDNRB is particularly interesting
Shrestha, Prashanta. « Streamlined Extract Preparation for E. coli-Based Cell-Free Protein Synthesis and Rapid Site-Specific Incorporation of Unnatural Amino Acids in Proteins ». BYU ScholarsArchive, 2012. https://scholarsarchive.byu.edu/etd/3917.
Texte intégralVOCCIA, DIEGO. « Electrochemical Genosensors for Tumor Biomarker Detection : the case of miRNAs ». Doctoral thesis, 2015. http://hdl.handle.net/2158/1003576.
Texte intégralChen, Chun-Cheng, et 陳俊丞. « PCR free detection of hepatitis B virus DNA using a nanostructured impedance biosensor ». Thesis, 2014. http://ndltd.ncl.edu.tw/handle/41461326880034226870.
Texte intégral國立中興大學
機械工程學系所
102
Chronic hepatitis B virus (HBV) infection is an important health burden because of its worldwide prevalence and potential adverse outcomes, including liver cirrhosis and hepatocellular carcinoma. Although the DNA level of HBV in the serum is an important biomarker associated with several important outcomes of HBV patients, it is not regularly monitored in clinical practice because of its high cost. So it is necessary to develop a highly sensitive and low-cost technique for effective detection of HBV concentration in blood. In this study, a PCR free technique for effective detection of HBV DNA obtained directly from clinical samples was presented. We use a sensitive nanostructured biosensor with a sensing electrode of gold nanoparticles (GNPs) uniformly deposited on a uniform nanohemisphere array. A specially designed single-strand gene fragment of the target genomic DNA of HBV based on the genome sequences of HBV was immobilized on the nanostructured electrode as the capture probe. Target HBV DNA obtained from clinical samples were hybridized to the sensing probes. Detection results illustrate two dynamic linear ranges, 10^2-10^3 and 10^3-10^5.1 copies/mL, having R^2 values of 0.801 and 0.996 could be obtained, respectively. The detection limit of the proposed sending scheme was measured to be 111 copies/mL. The total of 45 target samples, including 20 samples with HBV concentration being lower than 102 copies/mL and 25 samples with HBV concentration being in the range of 10^3-10^5.1 copies/mL. A detection limit of 186 copies/mL could be achieved. The concentration of these 45 HBV DNA samples was measured by the COBAS Ampliprep system. Comparing the measured results of the COBAS Ampliprep and our system, it was illustrated that the HBV DNA concentrations measured by the proposed method in this study had a high linear correlation with the COBAS Ampliprep, having R^2 values of 0.983 .The proposed sensing scheme is highly feasible for future clinical applications.
Govender, Kerushini. « Preliminary validation of Mycobacterium tuberculosis complex-specific PCR tests for the detection of M. bovis and M. tuberculosis in formalin-fixed, paraffin-embedded tissues of captive and free-ranging wildlife ». Diss., 2013. http://hdl.handle.net/2263/37366.
Texte intégralDissertation (MSc)--University of Pretoria, 2013.
gm2014
Veterinary Tropical Diseases
Unrestricted
Belšánová, Barbora. « Optimalizace metodiky pro stanovení volné nádorové DNA v plazmě a její klinické využití u pacientů s karcinomy kolorekta, plic a slinivky břišní ». Master's thesis, 2017. http://www.nusl.cz/ntk/nusl-368058.
Texte intégralKanzow, Philipp Clemens. « Zirkulierende Nukleinsäuren im zellfreien Plasma von LTx-Patienten als Frühmarker einer Schädigung des Spenderorgans ». Doctoral thesis, 2014. http://hdl.handle.net/11858/00-1735-0000-0022-5F66-1.
Texte intégralPazourková, Eva. « Analýza volných nukleových kyselin a její potenciální klinické využití ». Doctoral thesis, 2019. http://www.nusl.cz/ntk/nusl-396188.
Texte intégral