DB36T 1415-2021 猕猴桃雌雄种质分子身份证鉴定技术规程　　

DB36江西省市场监督管理局发布 猕猴桃雌雄种质分子身份证鉴定技术规程本文件规定了分子身份证鉴定猕猴桃（Actinidia）雌雄种质的术语和定义、原理、试剂与试药、2水浴锅水浴加热至65℃,预热3%裂解液、洗液，两种液体使用前均需加入2%体积的β-巯基乙醇）；硅胶干燥后的叶片样品需要进行的步骤。向2mL管子中加入1mL洗液，充分混匀，65℃水浴30硅胶干燥后的叶片样品需要进行的步骤。弃上清液，向2mL管子中加入1mL洗液，充分混匀，65取上清液（可0.80mL）于新的2mL管子中，加入等体积抽提液1，摇床混匀10min，随后12000取上清液（可0.60mL）于新的2mL管子中，加入等体积抽提液2，摇床混匀10min，随后12000-20℃沉淀30min以上，随后4℃,10000rrpm/min离心8min。此步骤94℃预变性3min，94℃变性30s，52℃~62℃退火30从银染液中取出凝胶，用双蒸水冲洗干净后，放入显影液中显影，待条带全部显出，转入到清水4利用同一样品的不同引物条带读数绘制成1/0矩阵，构建成该品种的特异性分子身份证，如附录6827100～150TCCAAAGCATGCTCAACTTC31CCAAGCTCATACACCCAATG57AGGTTGGCACTTGGAATAGG53TTCCTTAATCCAGGGTCTCGACACAACAGATTGCAGGAGGGAGTCGGGAGCATAATTGGTCGCATGGAAGGTCATAACAC100～150CGTCCTCTCTTCGCCATTTATGGTGTAAAGTCAAAAACAGCCAGAAGATTTCATTGTCCTCCCTAAGAGCCATAGCTTATTCACCG100～150AAGTAAAGCCATTGTCATTGCA1AACTAAGAAACGGGACCATTGTATGTGGATGTCCCAGAAAAG11CCCCTCATTGTAAAGCATCCTTGTCACGGGCTTAGAATCC15CCGTCTCAGCAGATGTCACTAT52GCAACTGCCCATAAGATTCC22TACACCTGATGAGATGGACGACCCTGTGGCTGATGTTGAAAGTTCGAGTTACCTAGCTACTCCGCCAAGGGAAGAAAATGTTGAACCGTAAGGTCATTTTTGCGAAAGGA017TACGCAGTTTACTTCCTCTTCATTCGGCACCACTTCTACCCATAGCCAACAACATCCTTGACGCCTCTGAACACA055TCCATTCCGCCCATCCTTA061ATCTCCATCATCTCCACCTT100～150CAACGCCATCATTGTCCCGAAGCAACTCGCTACTAAGAGCCTGTAACAACCCATT150～250TGCTATGGCACTCTATCCA103GAGCATACAGAGGGAAGAACTGGAGTGAAGGAGGGTA105CCTCATTCATTACCACCTCTTACACCGCTTCCGTCCATAA123CACCTGCTACACCGACATCCACAGCCGAAACCATACA124ATACCGTGGACTTCATTC150～250ACATACCTTCAACGCTTCA125GTTTATTACACGCCTTCGTGACAGCCCTCAACTACGA138CCCTATACCCACTCAATCATTCCACCTTCTTTCCATCA167ATGGCTTGACTGAGACTTG150～250A186CTCCCTCGTAGCAGTTCACTCCAGCGATTCCTTCACA191CCGCATCATAAGCCCAATCTTGTCCACCGCCCACTTCA193ATACCCATCATCGGACAC61150～250CTTCAAGCCCATTACCACCGACTAAAATCTACAAGCACGAAAACGAAATTAACACCATTAAGATTGATCGGCACG61GTTAATTTGATCGGGATGGGAGGAGCTTGAGCTGCTATATACACTTGAAGCGCCGC57AAGCAGCCATGTCGATACGTCCCCACACAACAACTCCTC10CATATGACCTGCACGTGCGGTTTGATCGGTCTTCGAAA57ATAAATGTGTGCCAGTGCGAGAGCGTAGAATGTGGCGG100～150CATTTGAAATGAGTCAAGCTGCAACGAAACAAAAGCCCTATTATCGCTTCAGATTCAGCCTCTGGTTCATCCCAATCCCTCATTCGATGGCTGACGAACAACATCTGTCACGATCACCTCCCCACTCTTCGCCTCTCTTTCA5727100～150TCCAAAGCATGCTCAACTTC53TTCCTTAATCCAGGGTCTCGACACAACAGATTGCAGGAGG100～150CGTCCTCTCTTCGCCATTTATGGTGTAAAGTCAAAAACAGCCAGAAGATTTCATTGTCCTCCCTAAGAGCCATAGCTTATTCACCG100～150AAGTAAAGCCATTGTCATTGCA1AACTAAGAAACGGGACCATTGTATGTGGATGTCCCAGAAAAG561CAGTCAGCCAAGGACFAAGG13ACTAACAGACAAAAACTGGGGGATGGAAGGAGATGGCGATG14TTGAGGGCTTGATGAAAACC61GACGACGACGGAGATTACG15CCGTCTCAGCAGATGTCACTAT52GCAACTGCCCATAAGATTCC22TACACCTGATGAGATGGACGACCCTGTGGCTGATGTTGAAAGTGTAAGGTCATTTTTGCGAAAGG12A017TACGCAGTTTACTTCCTCTTCATTCGGCACCACTTCTACCCATAGCCAACAACATCCTTGACGCCTCTGAACACA055TCCATTCCGCCCATCCTTA061ATCTCCATCATCTCCACCTT100～150CAACGCCATCATTGTCCCGAAGCAACTCGCTACTAAGAGCCTGTAACAACCCATT150～250TGCTATGGCACTCTATCCA105CCTCATTCATTACCACCTCTTACACCGCTTCCGTCCATAA123CACCTGCTACACCGACATCCACAGCCGAAACCATACA125GTTTATTACACGCCTTCGTGACAGCCCTCAACTACGA138CCCTATACCCACTCAATCATTCCACCTTCTTTCCATCA167ATGGCTTGACTGAGACTTG150～250A186CTCCCTCGTAGCAGTTCACTCCAGCGATTCCTTCACA191CCGCATCATAAGCCCAATCTTGTCCACCGCCCACTTCA193ATACCCATCATCGGACAC61150～250CTTCAAGCCCATTACCACESTAD01CGACTAAAATCTACAAGCACGAAAACGAAATTAACACCATTAAGATTGATCGGCACG61GTTAATTTGATCGGGATGGGAGGAGCTTGAGCTGCTATATACACTTGAAGCGCCGC57AAGCAGCCATGTCGATACGGGTTTGATCGGTCTTCGAAA57ATAAATGTGTGCCAGTGCGAGAGCGTAGAATGTGGCGG100～150CATTTGAAATGAGTCAAGCTGCACGTGACTTGGTTTTTGAAGG55CACTCCGATCAGCTCTCCTCAACGAAACAAAAGCCCTATTATCGCTTCAGATTCAGCCTCTGGTTCATCCCAATCCCTCATTCGATGGCTGACGAACAACATCTGTCACGATCACCTCCCCACTCTTCGCCTCTCTTTCA5714FOR-131200000101111011003400101111111100111110000100000111010105600001100111000001111110101078101111010000011000111010109100010100011000000111010010101110001011001213001110011000000011111011010141000111100000101010101111015100011110016000011110017000000101101110180000001011011110010111110110111111110011111101101010190000011010011100011111111100000001101101110000010110111010111111110001100001101010211000001010011112200111100110001001111101101023001001111112510101111010001111111100000100011110027101101110110110010111111110000111100001100001110111016100111001011010101020011100101101010103001110000010101110450011110100010160011110101101010107001110000010101110800111000001010111090011100000101011101011121001101010000010111310011010100000101114001011011010100011001500111001011010101016170010110110101000110018190010110110101000110000101101101010001100