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National Repository of Grey Literature 2 records found  Search took 0.00 seconds. 
Optimization of Real-Time PCR for detection of Klebsiella pneumoniae
 ELSTNEROVÁ, Šárka
The subject of this bachelor's thesis is the optimization of real-time PCR for the detection of the bacteria Klebsiella pneumoniae using primers from the publication Sun et al, 2010, for SYBR Green real-time PCR and end point PCR and primers designed by the supervisor of this thesis (Stehlíková, unpublished) for TaqMan real-time PCR. The selected primers were subsequently tested on pure culture samples of Klebsiella pneumoniae 4799 from the DSMZ collection and verified on 41 buccal swabs. Furthermore, assessment of specificity was accomplished using 26 bacterial DNA isolates. Assessment of sensitivity of PCR detection methods was accomplished depending on their routine use, assessment of their limits of detection and comparison.
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PCR based assay for detection garden pea lec gene
 Vráblík, Aleš ; Hodek, Jan ; Ovesná, Jaroslava
The method was developed and verified by staff of National reference laboratory for GMO identification and DNA fingerprinting suited for governmental control laboratories and private laboratories that analyze food and feed derived from various plant matrices. Method can be applied for quantification of GM pea in a sample, when reference gene is used as a standard. The method describe internal garden pea specific gene, lektin in this case, detection using PCR and real-time PCR. The general principle of the assay relies on lektin specific sequence presence that represents a species specific gene of garden pea. In many matrices DNA is damaged so amplification of short stretches of DNA is used. After amplification of target exploiting newly developer species specific gene primers PCR products are separated by electrophoresis in agarose gel. Resulting band is visualized by UV light and its position is compare with size standard. Alternatively, real-time PCR and ABI platform in combination with SYBR® Green can be used. Reaction parameters are described and specificity of reaction was verified. LOD as well as LOQ was defined.
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