RU2005104635A - METHOD FOR PROTECTING RT-PCR FROM CONTAMINATION BY AMPLIFICATION PRODUCTS BASED ON THE DESTRUCTIVE ACTION OF URAPYLES OF Glycosylase DNA - Google Patents

METHOD FOR PROTECTING RT-PCR FROM CONTAMINATION BY AMPLIFICATION PRODUCTS BASED ON THE DESTRUCTIVE ACTION OF URAPYLES OF Glycosylase DNA Download PDF

Info

Publication number
RU2005104635A
RU2005104635A RU2005104635/13A RU2005104635A RU2005104635A RU 2005104635 A RU2005104635 A RU 2005104635A RU 2005104635/13 A RU2005104635/13 A RU 2005104635/13A RU 2005104635 A RU2005104635 A RU 2005104635A RU 2005104635 A RU2005104635 A RU 2005104635A
Authority
RU
Russia
Prior art keywords
pcr
dna
contamination
protecting
dutp
Prior art date
Application number
RU2005104635/13A
Other languages
Russian (ru)
Other versions
RU2307167C2 (en
Inventor
Рамиль Ришадович Вафин (RU)
Рамиль Ришадович Вафин
Раиф Ришадович Вафин (RU)
Раиф Ришадович Вафин
Фани Файзрахмановна Замалиева (RU)
Фания Файзрахмановна Замалиева
Ирина Витальевна Пикалова (RU)
Ирина Витальевна Пикалова
Зенон Сташевски (RU)
Зенон Сташевски
Тахир Мунавирович Ахметов (RU)
Тахир Мунавирович Ахметов
Шамиль Касымович Шакиров (RU)
Шамиль Касымович Шакиров
Рустам Хамитович Равилов (RU)
Рустам Хамитович Равилов
Original Assignee
Рамиль Ришадович Вафин (RU)
Рамиль Ришадович Вафин
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Рамиль Ришадович Вафин (RU), Рамиль Ришадович Вафин filed Critical Рамиль Ришадович Вафин (RU)
Priority to RU2005104635/13A priority Critical patent/RU2307167C2/en
Publication of RU2005104635A publication Critical patent/RU2005104635A/en
Application granted granted Critical
Publication of RU2307167C2 publication Critical patent/RU2307167C2/en

Links

Landscapes

  • Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)

Claims (1)

Способ защиты ОТ-ПЦР от контаминации продуктами амплификации на основе разрушающего действия урацил ДНК гликозилазы, включающий подготовку пробы исследуемой рибонуклеиновой кислоты, внесение указанной пробы в реакционную смесь, состоящую из буферной системы, олигонуклеотидных праймеров, РНК и ДНК зависимых ДНК полимераз, урацил ДНК гликозилазы и dATP, dUTP, dCTP, dGTP, проведение обратнотранскриптазной полимеразной цепной реакции и анализ продуктов реакции методом гель-электрофореза на предмет выявления специфичного для определенного биологического объекта амплифицированного фрагмента ДНК, отличающийся тем, что в качестве РНК зависимой ДНК полимеразы применяют обратную транскриптазу M-MuLV, а ОТ-ПЦР проводят в варианте "1 пробирка, 2 этапа" путем комбинированного использования dTTP/dUTP, что достигается внесением на первом этапе обратной транскрипции dATP, dTTP, dCTP, dGTP, а на втором этапе полимеразной цепной реакции dATP, dUTP, dCTP, dGTP.A method for protecting RT-PCR from contamination with amplification products based on the destructive action of uracil DNA glycosylase, including preparing a sample of the studied ribonucleic acid, introducing said sample into a reaction mixture consisting of a buffer system, oligonucleotide primers, RNA and DNA-dependent DNA polymerases, uracil DNA glycosylase and dATP, dUTP, dCTP, dGTP, reverse transcriptase polymerase chain reaction and analysis of reaction products by gel electrophoresis to identify specific for a particular biologic object of the amplified DNA fragment, characterized in that the reverse transcriptase M-MuLV is used as the RNA-dependent DNA polymerase, and RT-PCR is carried out in the "1 tube, 2 steps" variant by combined use of dTTP / dUTP, which is achieved by introducing in the first step reverse transcription dATP, dTTP, dCTP, dGTP, and in the second stage of the polymerase chain reaction dATP, dUTP, dCTP, dGTP.
RU2005104635/13A 2005-02-21 2005-02-21 Method for protection of rt-pcr from contamination by amplification products based on destructive action of uracyl dna glycosylase RU2307167C2 (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
RU2005104635/13A RU2307167C2 (en) 2005-02-21 2005-02-21 Method for protection of rt-pcr from contamination by amplification products based on destructive action of uracyl dna glycosylase

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
RU2005104635/13A RU2307167C2 (en) 2005-02-21 2005-02-21 Method for protection of rt-pcr from contamination by amplification products based on destructive action of uracyl dna glycosylase

Publications (2)

Publication Number Publication Date
RU2005104635A true RU2005104635A (en) 2006-08-10
RU2307167C2 RU2307167C2 (en) 2007-09-27

Family

ID=37058908

Family Applications (1)

Application Number Title Priority Date Filing Date
RU2005104635/13A RU2307167C2 (en) 2005-02-21 2005-02-21 Method for protection of rt-pcr from contamination by amplification products based on destructive action of uracyl dna glycosylase

Country Status (1)

Country Link
RU (1) RU2307167C2 (en)

Families Citing this family (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
RU2717975C2 (en) * 2018-08-06 2020-03-27 Общество с ограниченной ответственностью "ОНКОДИАГНОСТИКА АТЛАС" Method for estimating cross-contamination of samples in ngs-data on analysis of high-polymorphic loci

Also Published As

Publication number Publication date
RU2307167C2 (en) 2007-09-27

Similar Documents

Publication Publication Date Title
JP2010004884A5 (en)
IL102896A (en) Process, kit and nucleic acid reagents for hepatitis c detection
EP1645640A3 (en) Methods for amplifying and analyzing nucleic acids
WO2009042291A3 (en) Accelerated cascade amplification (aca) of nucleic acids comprising strand and sequence specific dna nicking
JP2013500705A (en) Nucleic acid normalized quantification method based on ligation
JP2008136451A (en) Method for amplifying nucleic acid
JP2016527896A5 (en)
RU2006118406A (en) METHOD FOR DETECTION OF BACTERIA OF THE GENUS YERSINIA AND DIFFERENTIATION OF PATHOGENIC FOR HUMAN SPECIES OF JERSIN BY THE METHOD OF POLYMERASE CHAIN REACTION (PCR), OLIGONUCLEOTIDE PRIMERS AND TESTING
JP2006166907A5 (en)
CN105986043A (en) Method for quickly detecting nucleic acid of H5 subtype highly pathogenic avian influenza virus
CN105986042A (en) Method for quickly detecting nucleic acid of H9 subtype avian influenza virus
CN105986044A (en) Universal method for quickly detecting nucleic acid of avian influenza virus
CN117070669A (en) Novel coronavirus whole genome capture kit
RU2005104635A (en) METHOD FOR PROTECTING RT-PCR FROM CONTAMINATION BY AMPLIFICATION PRODUCTS BASED ON THE DESTRUCTIVE ACTION OF URAPYLES OF Glycosylase DNA
ATE526418T1 (en) IMPROVED COMPOSITIONS FOR IN VITRO AMPLIFICATION OF NUCLEIC ACIDS
Aifat et al. Ancient DNA analyses of museum specimens from selected Presbytis (Primate: Colobinae) based on partial Cyt b sequences
EP3533884A1 (en) Dna sequences to assess contamination in dna sequencing
DE602004030036D1 (en) Multiplex analysis of nucleic acids for the determination of human papillomavirus genotypes
CN106011307A (en) Nucleic acid rapid detection method for H7 subtype avian influenza virus
US20030104421A1 (en) Methods and compositions for nucleic acid amplification
Bakar et al. Optimisation of polymerase chain reaction conditions to amplify D-loop region in the Malaysian mousedeer genomic DNA
Kitade et al. Effect of DMSO on PCR of Porphyra yezoensis (Rhodophyta) gene
JP2007000040A (en) Method for detecting b-type hepatitis virus
RU2526499C1 (en) SET OF SYNTHETIC OLIGONUCLEOTIDES FOR SPECIES IDENTIFICATION OF ROSEROOT (Rhodiola quadrifida (Pall) Fisch et Mey)
US20240301517A1 (en) A microfluidic pipeline for isolation and analysis of single viruses

Legal Events

Date Code Title Description
MM4A The patent is invalid due to non-payment of fees

Effective date: 20090222