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LY/T 1772-2008: Molecular identification method for varieties of poplar. Amplified fragment length polymorphism(AFLP)
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Molecular identification method for varieties of poplar. Amplified fragment length polymorphism(AFLP)
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LY/T 1772-2008
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Basic data | Standard ID | LY/T 1772-2008 (LY/T1772-2008) | | Description (Translated English) | Molecular identification method for varieties of poplar. Amplified fragment length polymorphism(AFLP) | | Sector / Industry | Forestry Industry Standard (Recommended) | | Classification of Chinese Standard | B61 | | Classification of International Standard | 65.020 | | Word Count Estimation | 12,122 | | Date of Issue | 2008-09-03 | | Date of Implementation | 2008-12-01 | | Regulation (derived from) | Industry standard filing Notice No. 10 of 2008 | | Issuing agency(ies) | State Forestry Administration | | Summary | This standard specifies the poplar fresh tissue (leaves, buds, etc.) for the material, the use of amplified fragment length polymorphism DNA technique (Amplified Fragment Length Polymorphism, referred AFLP) molecular identification of poplar varieties experimental method. This standard applies to molecular identification of poplar species. | LY/T 1772-2008: Molecular identification method for varieties of poplar. Amplified fragment length polymorphism(AFLP)
---This is a DRAFT version for illustration, not a final translation. Full copy of true-PDF in English version (including equations, symbols, images, flow-chart, tables, and figures etc.) will be manually/carefully translated upon your order.
Molecular identification method for varieties of poplar.Amplified fragment length polymorphism (AFLP)
ICS 65.020
B61
People's Republic of China Forestry Industry Standard
Experimental method for molecular identification of poplar varieties
DNA amplified fragment length polymorphism (AFLP)
Released on.2008-09-03
2008-12-01 implementation
State Forestry Administration released
Foreword
Appendix A of this standard is an informative annex.
This standard was proposed by the State Forestry Administration.
This standard is under the jurisdiction of the National Forest Seed Standardization Technical Committee.
This standard was drafted. Forestry Research Institute of Chinese Academy of Forestry, Institute of Genetics and Developmental Biology, Chinese Academy of Sciences and National Forest
Bureau of Northern Forest Seed Testing Center.
The main drafters of this standard. Li Jinhua, Zhang Weiwen, Wang Bin, Song Hongzhu, Zhou Chunjiang, Lu Mengzhu, Li Qingmei, Niu Zhengtian.
Experimental method for molecular identification of poplar varieties
DNA amplified fragment length polymorphism (AFLP)
1 Scope
This standard stipulates that the fresh tissue of poplar (leaf, bud, etc.) is used as the material, and the amplified fragment length polymorphism DNA method (Amplified)
FragmentLengthPolymorphism (AFLP) is an experimental method for molecular identification of poplar varieties.
This standard applies to the molecular identification of poplar varieties.
2 Principle
The primers matched by the adjacent cleavage sites are amplified, and finally the specific fragments are separated by denaturing polyacrylamide gel electrophoresis.
Determine whether the tested variety and the control variety are the same by comparing the difference in DNA fingerprint band data between the tested variety and the control variety.
Variety.
3 Instruments and equipment
3.1 General laboratory equipment.
3.2 Gradient PCR Amplifier.
3.3 Agarose gel electrophoresis system.
3.4 Denatured polyacrylamide gel electrophoresis system.
3.5 UV transilluminator.
3.6 Gel imaging system or camera system.
3.7 Heavy distilled water meter.
4 reagents and solutions
Analytically pure reagents were used in the analysis unless otherwise stated. The prepared solution is used after autoclaving. Water used in the experiment
All are sterile ultrapure water.
Store at -20 °C.
4.2 T犪狇DNA polymerase (1U/μL) and its 10×PCR reaction buffer
10×PCR reaction buffer containing.200 mmol/L Tris-HCl (pH 8.4), 15 mmol/L magnesium chloride (MgCl 2 ) and
500 mmol/L potassium chloride (KCl), stored at -20 °C.
4.3 T4 DNA ligase (1U/μL) and its 10× ligation reaction buffer
10× reaction buffer containing 350 mmol/LTris-HCl (pH 7.6), 50 mmol/L MgCl 2 and 500 mmol/L KCl and
5 mmol/L β-mercaptoethanol; stored at -20 °C.
Store at -20 °C.
λDNA and DNAMarker DL2000 were stored at -20 °C.
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