求高人帮忙翻译下文章 生物类的

来源:百度知道 编辑:UC知道 时间:2024/06/17 19:17:33
Gene expression and real-time RT-PCR analysis
For qRT-PCR analysis, total RNAs were isolated by using the SV total RNA isolation kit (Promega). SuperScriptTM III reverse transcriptase (Invitrogen) was used to synthesize the first-strand cDNA with oligo dT primer (or random hexamers from Thermo Scientific, UK for the analysis of FLC expression in FLC transgenic plants that carry endogenous FLC and FLC transgene) and 1 μg of total RNA at 50 °C for 1 h. Quantitative PCR was then performed with Platinum SYBR Green qRT-PCR kit (Invitrogen) on Applied Biosystems 7500 Real-Time PCR machine. The thermal cycling program was 50 °C for 10 min and 95 °C for 10 min, followed by 40 cycles of 95 °C for 30 s, 56 °C for 30 s, 72 °C for 1 min and a one-cycle dissociation stage at 95 °C for 15 s, 60 °C for 1 min, and 95 °C for 15 s. The primer pairs used in qRT-PCR were FT, 5′-GAGACCCTCTTATAGTAAGCA-3′ and 5′-CTTCCTCCGCAGCCACTCA-3′; SOC1, 5′-AATATGCAAGATACCATAGATCGT-3′ and 5′-TTCTTGAAGAACAAGGTAACCCAA

基因表达和实时RT - PCR分析
为qRT - PCR分析,总RNA分离使用队总RNA提取试剂盒(盒) 。 SuperScriptTM三逆转录酶( Invitrogen公司)被用来合成第一链cDNA与寡核苷酸引物(或随机hexamers由热科学,英国的分析刚果解放阵线的表达联合体从事转基因植物内源基因刚果解放阵线和刚果解放运动)和1微克总RNA ,在50 ° C的1小时定量聚合酶链反应,然后用铂荧光绿色qRT - PCR检测试剂盒( Invitrogen公司)的应用生物系统公司7500实时PCR机器。在热循环程序为50 ° C的10分钟和95 ° C的10分,其次是40个周期为95 ° C的30秒, 56 ° C的30秒, 72 ° C的1分和一个周期解舞台上95 ° C的15章, 60 ° C时为1分,和95 ° C的15号引物对用于qRT - PCR检测了红外, 5' - GAGACCCTCTTATAGTAAGCA - 3 '和5' - CTTCCTCCGCAGCCACTCA - 3 ' ; SOC1 , 5' - AATATGCAAGATACCATAGATCGT - 3 '和5' - TTCTTGAAGAACAAGGTAACCCAA - 3 ' ;二氧化碳, 5' - ACGCCATCAGCGAGTTCC - 3 '和5' - AAATGTATGCGTTATGGTTAATGG - 3 ' ;联合体, 5' - AGTAGCCGACAAGTCACCTT - 3 '和5' - GAGAGTCACCGGAAGATT - 3 ' ;激光器, 5' - AACAGCAACAATATATGCAAC - 3 '和5' - ATATCCTGGATTGCTACTCAT - 3 ' ; UBQ10 , 5' - AGGTACAGCGAGAGAAAGTAGCA - 3 '和5' - TAGGCATAGCGGCGAGGCGT - 3 ' 。数据计算了从三个生物样品和生物样品每检查一式三份。
为组织特异性表达分析,不同的组织类型是收获的植物和立即冻结液氮,包括5天前