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Czech to English: Diagnostic kit for detection of the minimal residual disease of pancreatic carcinoma General field: Science Detailed field: Medical: Instruments
Source text - Czech Diagnostický kit pro detekci minimální reziduální choroby u karcinomu pankreatu
Polymerázová řetězová reakce v reálném čase (real-time PCR) je enzymatická reakce, která je umožněna primery, specifickou fluorescenčně značenou sondou a termostabilní DNA-polymerázou, kdy průběh reakce je snímán termocyklérem v reálném čase na základě změn fluorescence. Po přidání standardů k reakci a vytvoření standardizační křivky lze hodnotit množství specifické vyšetřované sekvence ve vzorku. Ke každé polymeráze je dodáván reakční pufr o vhodném složení. Optimální koncentrace hořečnatých iontů by měla být stanovena pro každý nový primerový pár, novou polymerázu a nový termocyklér. Výhodné je použití DNA-polymerázy s tzv. „HotStartem“, pro jejíž aktivaci je nutné 10 až 15 minutové zahřívání na 90-95°C. Primery jsou krátké oligonukleotidy (o délce cca 20bp), které vymezují amplifikovaný úsek DNA, specifická hydrolyzační TaqMan sonda je krátký oligonukleotid značený fluorescenční barvou a zhášečem. Teplota nasedání primerů je orientačně vypočítána z nukleotidového složení a měla by být následně experimentálně ověřena při optimalizaci amplifikačního cyklu.
Postup:
Chemikálie uchováváme při – 18°C až – 30°C. Před začátkem práce rozmrazíme na ledové tříšti CEA sense 0,015mM, CEA antisense 0,015mM, CEA probe 1,25цM, EGFR sense 0,005mM, EGFR antisense 0,005mM, EGFR probe 1,25цM, hTERT sense 0,005mM, hTERT antisense 0,005mM, hTERT probe 1,25цM, dNTP 10mM mix, Water. Připravíme vhodnou HotStart Taq polymerázu s patřičným pufrem a MgCl2. Po rozpuštění vše krátce zvortexujeme a stočíme. HotStart Taq polymeráza se přidává přímo z mrazáku (pouze krátce stočit). Zapneme real-time termocyklér.
Translation - English Diagnostic kit for detection of the minimal residual disease of pancreatic carcinoma
Real time polymerase chain reaction (real-time PCR) is an enzymatic reaction, facilitated by primers, a specific fluorescently labeled probe and a thermostable DNA polymerase. The reaction progress is monitored in real time by thermocycler based on changes in fluorescence. By adding standards to the reaction and plotting a standard curve, a quantity of the specific sequence of interest in the sample can be evaluated. A reaction buffer of a suitable composition is provided for each polymerase. The optimal concentration of magnesium ions should be established for each new primer pair, a new polymerase and a new thermocycler. The use of the so-called “HotStart” DNA polymerase which requires 10-15 minutes heating to 90-95ºC for its activation is beneficial. Primers are short oligonucleotides (about 20 bp long) that delineate the amplified region of the DNA. The specific hydrolysis TaqMan probe is a short fluorescently labeled oligonucleotide with a quencher. In first approximation, the primer annealing temperature is calculated from the nucleotide composition and should be further verified experimentally during the optimization of the amplification cycle.
Procedure:
Store reagents at -18º C to – 30º C. Prior to commencement of work thaw the following reagents on crushed ice: CEA sense 0.015mM, CEA antisense 0.015mM, CEA probe 1.25 µM , EGFR sense 0.005mM, EGFR antisense 0.005mM, EGFR probe 1.25 µM, hTERT sense 0.005mM, hTERT antisense 0.005mM, hTERT probe 1.25 µM, dNTP 10mM mix, water. Prepare the suitable HotStart polymerase with the corresponding buffer and MgCl2. After dissolving, vortex the solution shortly and spin down. HotStart Taq polymerase is added directly from the freezer (just spin down shortly). Turn on the real-time thermocycler.
Until recently, I had worked in a biomedical research laboratory at a Canadian university. I authored and co-authored scientific manuscripts on atherosclerosis, kidney disease, lung disease and immunohistochemistry in peer-reviewed science journals. I have good understanding of scientific and medical terminology and concepts which leads to correct translation.
I had lived in Czechoslovakia for 33 years, graduated as Dipl. Engineer (equivalent to MSc) from the School of Chemical Technology in 1974, passed the State exam in English at the School of Languages in Prague, in 1968, earned a PhD degree in Biochemistry from the Czech Academy of Science in 1996.
I have been translating medical, pharmaceutical, scientific and clinical trial documents for 4 years. I have completed 53 translation and editing projects.
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Member since Sep '09
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