首页>
外国专利>
Process for simultaneous preparation of sex specific and gender-neutral semisynthetic amplicons useful for sex determination
Process for simultaneous preparation of sex specific and gender-neutral semisynthetic amplicons useful for sex determination
展开▼
机译：同时制备可用于性别确定的性别特异性和性别中性半合成扩增子的方法
展开▼
页面导航
摘要
著录项
相似文献
摘要
An improved process for simultaneous preparation of sex specific and gender-neutral semisynthetic amplicons obtained by amplifying sequences of synthetic oligonucleotide primers identified as SEQ ID NOS:4 to 7 (5'GGATCCCTATlAGTG 3'; 5'GGATCCCITTTGCACTC 3'; 5CGAAATCGGTAGACGATACG3' and 5'GGGGATAGAGGGACTTGAAC 3') useful for sex determination, said process comprising isolating nucleic acids from any part of a papaya plant by conventional methods, amplifying the said nucleic acids in a conventional Random Amplification of polymorphic DNA Polymerase Chain Reaction (RAPD-PCR), resolving the amplified products by conventional electrophoresis method, eluting the sex specific, double stranded amplified product from the gel piece by known methods, cloning the said product in a known vector by conventional methods, sequencing the said cloned product by known methods, synthesizing the single stranded chains of synthetic oligonucleotides by known method based on the said sequence data, amplifying the said nucleic acid in a Conventional Sequence Tagged Site Polymerase Chain Reaction by using synthetic oligonucleotides as primers to get sex specific and gender-neutral semisynthetic amplicons simultaneously.
展开▼
机译：一种同时制备性别特异性和性别中性的半合成扩增子的改进方法，该扩增子通过扩增鉴定为SEQ ID NOS：4至7的合成寡核苷酸引物的序列而获得（5'GGATCCCTATlAGTG 3'; 5'GGATCCCITTTGCACTC 3'; 5CGAAATCGGTAGACGATACG3'和5'用于性别确定的GGGGATAGAGGGACTTGAAC 3'），所述方法包括通过常规方法从木瓜植物的任何部分中分离核酸，在常规的多态性DNA聚合酶链反应（RAPD-PCR）的随机扩增中扩增所述核酸，通过常规电泳方法扩增产物，通过已知方法从凝胶片段上洗脱性别特异性的双链扩增产物，通过常规方法将所述产物克隆到已知载体中，通过已知方法对所述克隆产物进行测序，合成单链基于所述序列数据，通过已知方法合成寡核苷酸，扩增在常规序列标记位点聚合酶链反应中，通过使用合成寡核苷酸作为引物来获得所述核酸，以同时获得性别特异性和性别中性的半合成扩增子。
展开▼
