RU2015105268A - A method for identifying raspberry varieties based on RAPD markers - Google Patents

A method for identifying raspberry varieties based on RAPD markers Download PDF

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RU2015105268A
RU2015105268A RU2015105268A RU2015105268A RU2015105268A RU 2015105268 A RU2015105268 A RU 2015105268A RU 2015105268 A RU2015105268 A RU 2015105268A RU 2015105268 A RU2015105268 A RU 2015105268A RU 2015105268 A RU2015105268 A RU 2015105268A
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oligo
variety
dna
amplification products
raspberry
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RU2015105268A
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RU2620967C2 (en
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Вадим Георгиевич Лебедев
Михаил Викторович Филиппов
Анна Борисовна Азарова
Дмитрий Ильич Каган
Константин Александрович Шестибратов
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Общество с ограниченной ответственностью Научно-производственное предприятие "МИКРОКЛОН" (ООО НПП "МИКРОКЛОН")
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    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
    • C12Q1/6844—Nucleic acid amplification reactions
    • C12Q1/686—Polymerase chain reaction [PCR]

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  • Chemical & Material Sciences (AREA)
  • Organic Chemistry (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Zoology (AREA)
  • Wood Science & Technology (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
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  • General Health & Medical Sciences (AREA)
  • Genetics & Genomics (AREA)
  • Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)

Abstract

1. Способ идентификации сортов малины на основе RAPD-маркеров, включающий отбор растительного материала, выделение ДНК из растительного материала, амплификацию ДНК в реакционной смеси с последовательным участием нескольких рандомных праймеров, разделение продуктов амплификации с помощью электрофореза, визуализацию продуктов амплификации под ультрафиолетовым светом и выявление среди продуктов амплификации присущих только для данного сорта профилей, отличающийся тем, что амплификацию ДНК проводят с последовательным участием следующих рандомных праймеров:Oligo 4 - 5'-CAAACGGCAC-3';Oligo 6 - 5'-CACGGCGAGT-3';Oligo 29 - 5'-AAGAGCCCGT-3';Oligo 94 - 5'-GGACGGGTGC-3';Oligo 98 - 5'-GGGTAACGCC-3';Oligo 105 - 5'-AATCGGGCTG-3'.2. Способ по п. 1, отличающийся тем, что для каждого рандомного праймера обеспечивают индивидуальную температуру отжига (°С):Oligo 4 - 34,5;Oligo 6 - 38,3;Oligo 29 - 35,1;Oligo 94 - 41,5;Oligo 98 - 36,7;Oligo 105 - 37,3.3. Способ по п. 1, отличающийся тем, что выделение ДНК осуществляют из материала, взятого от не менее десяти растений каждого сорта, выращенных в поле или защищенном грунте или не менее двадцати растений каждого сорта, выращенных в условиях in vitro.4. Способ по п. 1, отличающийся тем, что на основе присущих только для данного сорта профилей составляют генетический паспорт сорта, включающий сочетание локусов и путем сравнения с которым проводят идентификацию сорта малины.1. A method for identifying raspberry varieties based on RAPD markers, including selecting plant material, isolating DNA from plant material, amplifying the DNA in a reaction mixture with sequential participation of several random primers, separating amplification products by electrophoresis, visualizing amplification products under ultraviolet light, and detecting among the amplification products inherent only for this variety of profiles, characterized in that the DNA amplification is carried out with the sequential participation of the following random primers: Oligo 4 - 5'-CAAACGGCAC-3 '; Oligo 6 - 5'-CACGGCGAGT-3'; Oligo 29 - 5'-AAGAGCCCGT-3 '; Oligo 94 - 5'-GGACGGGTGC-3'; Oligo 98 - 5'-GGGTAACGCC-3 '; Oligo 105 - 5'-AATCGGGCTG-3'.2. The method according to claim 1, characterized in that for each randomized primer an individual annealing temperature (° C) is provided: Oligo 4 - 34.5; Oligo 6 - 38.3; Oligo 29 - 35.1; Oligo 94 - 41.5 ; Oligo 98 - 36.7; Oligo 105 - 37.3.3. The method according to claim 1, characterized in that the DNA extraction is carried out from material taken from at least ten plants of each cultivar grown in a field or sheltered ground or at least twenty plants of each cultivar grown in vitro. The method according to claim 1, characterized in that on the basis of profiles inherent only for a given variety, a genetic passport is made for the variety, including a combination of loci and by comparison with which the raspberry variety is identified.

Claims (4)

1. Способ идентификации сортов малины на основе RAPD-маркеров, включающий отбор растительного материала, выделение ДНК из растительного материала, амплификацию ДНК в реакционной смеси с последовательным участием нескольких рандомных праймеров, разделение продуктов амплификации с помощью электрофореза, визуализацию продуктов амплификации под ультрафиолетовым светом и выявление среди продуктов амплификации присущих только для данного сорта профилей, отличающийся тем, что амплификацию ДНК проводят с последовательным участием следующих рандомных праймеров:1. A method for identifying raspberry varieties based on RAPD markers, including selecting plant material, isolating DNA from plant material, amplifying the DNA in a reaction mixture with sequential participation of several random primers, separating amplification products by electrophoresis, visualizing amplification products under ultraviolet light, and detecting among the amplification products inherent only for this variety of profiles, characterized in that the DNA amplification is carried out with the sequential participation of the following random primers: Oligo 4 - 5'-CAAACGGCAC-3';Oligo 4-5'-CAAACGGCAC-3 '; Oligo 6 - 5'-CACGGCGAGT-3';Oligo 6 - 5'-CACGGCGAGT-3 '; Oligo 29 - 5'-AAGAGCCCGT-3';Oligo 29-5'-AAGAGCCCGT-3 '; Oligo 94 - 5'-GGACGGGTGC-3';Oligo 94 - 5'-GGACGGGTGC-3 '; Oligo 98 - 5'-GGGTAACGCC-3';Oligo 98-5'-GGGTAACGCC-3 '; Oligo 105 - 5'-AATCGGGCTG-3'.Oligo 105 - 5'-AATCGGGCTG-3 '. 2. Способ по п. 1, отличающийся тем, что для каждого рандомного праймера обеспечивают индивидуальную температуру отжига (°С):2. The method according to p. 1, characterized in that for each randomized primer provide an individual annealing temperature (° C): Oligo 4 - 34,5;Oligo 4 - 34.5; Oligo 6 - 38,3;Oligo 6 - 38.3; Oligo 29 - 35,1;Oligo 29 - 35.1; Oligo 94 - 41,5;Oligo 94 41.5; Oligo 98 - 36,7;Oligo 98 - 36.7; Oligo 105 - 37,3.Oligo 105 - 37.3. 3. Способ по п. 1, отличающийся тем, что выделение ДНК осуществляют из материала, взятого от не менее десяти растений каждого сорта, выращенных в поле или защищенном грунте или не менее двадцати растений каждого сорта, выращенных в условиях in vitro.3. The method according to p. 1, characterized in that the extraction of DNA is carried out from material taken from at least ten plants of each cultivar grown in a field or protected ground or at least twenty plants of each cultivar grown in vitro. 4. Способ по п. 1, отличающийся тем, что на основе присущих только для данного сорта профилей составляют генетический паспорт сорта, включающий сочетание локусов и путем сравнения с которым проводят идентификацию сорта малины. 4. The method according to p. 1, characterized in that on the basis of the profiles inherent only for a given variety, a genetic passport is made for the variety, including a combination of loci and by comparison with which the raspberry variety is identified.
RU2015105268A 2015-02-17 2015-02-17 Method of identifying raspberry varieties based on rapd-markers RU2620967C2 (en)

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Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
RU2671678C2 (en) * 2017-04-14 2018-11-06 Общество с ограниченной ответственностью "ФИЛАГР" (ООО "ФИЛАГР") Method of genetic certification of raspberry selection achievements based on rapd-markers

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* Cited by examiner, † Cited by third party
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RU2732922C2 (en) * 2018-12-25 2020-09-24 Федеральное государственное бюджетное образовательное учреждение высшего образования "Пущинский государственный естественно-научный институт" (ПущГЕНИ) Method for molecular labelling, based on microsatellite loci, intended for genetic certification of plant variety of rubus genus plants

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RU2244416C2 (en) * 2002-02-06 2005-01-20 Всероссийский научно-исследовательский институт кормов им. В.Р. Вильямса Method for marking selection in achievement of meadow clover based on rapd-markers

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
RU2671678C2 (en) * 2017-04-14 2018-11-06 Общество с ограниченной ответственностью "ФИЛАГР" (ООО "ФИЛАГР") Method of genetic certification of raspberry selection achievements based on rapd-markers

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