【技术实现步骤摘要】
一种高通量基因表达谱检测试剂盒
本专利技术涉及一种高通量基因表达谱检测试剂盒及使用方法，适用于任何具有细胞基因表达水平检测需求的样本，涵盖任何真核生物的细胞。属于生物医学领域。
技术介绍
核酸是遗传信息的载体，是最重要的生物信息分子，是分子生物学研究的主要对象。随着精准医疗计划的进展，高通量基因测序技术将在以后的疾病精确诊断、疾病亚型分类、精准监控疾病发生发展、精准指导用药、特异疾病相关基因组数据库建立等方面发挥越来越关键作用。高通量测序的实现离不开样本的收集及测序前样本准备工作。尽管样本从收集到建库的成本显著降低，然而一旦涉及高通量、大量样本的建库，成本依然较高。目前商品化的转录组文库构建试剂盒几乎为进口试剂盒，价格昂贵，且试剂盒中各种成分相对独立，操作繁琐。因此，本专利技术提供了一种高通量基因表达谱检测试剂盒及使用方法，将整个样本建库前的所有处理步骤模块化，实现了样本处理的简易化、低起始量化、低成本化、高通量化。
技术实现思路
本专利技术的试剂盒主要用于高通量的基因表达谱的检测，该检测试剂盒提供了从样本收集到mRNA测序前的所有实验步骤所需的试剂及实验方法；一共可分为四个部分：1.逆转录反应模块，核酸纯化模块，双链cDNA合成模块，测序文库构建模块。本专利技术采用的技术方案为：一种高通量基因表达谱检测试剂盒，该试剂盒包括逆转录反应模块、核酸纯化模块、双链cDNA合成模块、测序文库构建模块；该逆转录反应模块，包括细胞裂解液、逆转录Mix、Barcode板(即逆转录引物板)...
【技术保护点】
1.一种高通量基因表达谱检测试剂盒，其特征在于：该试剂盒包括逆转录反应模块、核酸纯化模块、双链cDNA合成模块、测序文库构建模块；/n该逆转录反应模块，包括细胞裂解液、逆转录Mix、Barcode板、备用96孔板；其中：/n细胞裂解液：用pBS配置的0.3％Triton-100；/n逆转录Mix：0.5μl dNTP,0.25μl逆转录酶,1μl 5×逆转录酶Buffer[250mM Tris-HCl(pH 8.3),375mM KCl,15mM MgCl
【技术特征摘要】
1.一种高通量基因表达谱检测试剂盒，其特征在于：该试剂盒包括逆转录反应模块、核酸纯化模块、双链cDNA合成模块、测序文库构建模块；
该逆转录反应模块，包括细胞裂解液、逆转录Mix、Barcode板、备用96孔板；其中：
细胞裂解液：用pBS配置的0.3％Triton-100；
逆转录Mix：0.5μldNTP,0.25μl逆转录酶,1μl5×逆转录酶Buffer[250mMTris-HCl(pH8.3),375mMKCl,15mMMgCl2],0.125μlDTT,0.125μlRNA酶抑制剂,1μlbetaine(5M),0.045μlMgCl2；
Barcode板：含96种不同标签的逆转录引物(浓度为20μM)：(5’—3’)
ACACTCTTTCCCTACACGACGCTCTTCCGATCTCTCACATTTTTTTTTTTTTTTTTTTTTTTTTTTTTT(SEQIDNO:1)
ACACTCTTTCCCTACACGACGCTCTTCCGATCTTACTGCTTTTTTTTTTTTTTTTTTTTTTTTTTTTTT(SEQIDNO:2)
ACACTCTTTCCCTACACGACGCTCTTCCGATCTATCAGCTTTTTTTTTTTTTTTTTTTTTTTTTTTTTT(SEQIDNO:3)
ACACTCTTTCCCTACACGACGCTCTTCCGATCTCGACTATTTTTTTTTTTTTTTTTTTTTTTTTTTTTT(SEQIDNO:4)
ACACTCTTTCCCTACACGACGCTCTTCCGATCTCGTCATTTTTTTTTTTTTTTTTTTTTTTTTTTTTTT(SEQIDNO:5)
ACACTCTTTCCCTACACGACGCTCTTCCGATCTCTCGATTTTTTTTTTTTTTTTTTTTTTTTTTTTTTT(SEQIDNO:6)
ACACTCTTTCCCTACACGACGCTCTTCCGATCTTAGCACTTTTTTTTTTTTTTTTTTTTTTTTTTTTTT(SEQIDNO:7)
ACACTCTTTCCCTACACGACGCTCTTCCGATCTTCTAGCTTTTTTTTTTTTTTTTTTTTTTTTTTTTTT(SEQIDNO:8)
ACACTCTTTCCCTACACGACGCTCTTCCGATCTCACGTATTTTTTTTTTTTTTTTTTTTTTTTTTTTTT(SEQIDNO:9)
ACACTCTTTCCCTACACGACGCTCTTCCGATCTATGCTCTTTTTTTTTTTTTTTTTTTTTTTTTTTTTT(SEQIDNO:10)
ACACTCTTTCCCTACACGACGCTCTTCCGATCTGCTACATTTTTTTTTTTTTTTTTTTTTTTTTTTTTT(SEQIDNO:11)
ACACTCTTTCCCTACACGACGCTCTTCCGATCTGCATCTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTT(SEQIDNO:12)
ACACTCTTTCCCTACACGACGCTCTTCCGATCTACATGCTTTTTTTTTTTTTTTTTTTTTTTTTTTTTT(SEQIDNO:13)
ACACTCTTTCCCTACACGACGCTCTTCCGATCTACGTCATTTTTTTTTTTTTTTTTTTTTTTTTTTTTT(SEQIDNO:14)
ACACTCTTTCCCTACACGACGCTCTTCCGATCTCATGCTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTT(SEQIDNO:15)
ACACTCTTTCCCTACACGACGCTCTTCCGATCTCTAGTCTTTTTTTTTTTTTTTTTTTTTTTTTTTTTT(SEQIDNO:16)
ACACTCTTTCCCTACACGACGCTCTTCCGATCTTGCATCTTTTTTTTTTTTTTTTTTTTTTTTTTTTTT(SEQIDNO:17)
ACACTCTTTCCCTACACGACGCTCTTCCGATCTCATCTGTTTTTTTTTTTTTTTTTTTTTTTTTTTTTT(SEQIDNO:18)
ACACTCTTTCCCTACACGACGCTCTTCCGATCTCAGATCTTTTTTTTTTTTTTTTTTTTTTTTTTTTTT(SEQIDNO:19)
ACACTCTTTCCCTACACGACGCTCTTCCGATCTAGCACTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTT(SEQIDNO:20)
ACACTCTTTCCCTACACGACGCTCTTCCGATCTACTCGATTTTTTTTTTTTTTTTTTTTTTTTTTTTTT(SEQIDNO:21)
ACACTCTTTCCCTACACGACGCTCTTCCGATCTTCACGTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTT(SEQIDNO:22)
ACACTCTTTCCCTACACGACGCTCTTCCGATCTGTCTACTTTTTTTTTTTTTTTTTTTTTTTTTTTTTT(SEQIDNO:23)
ACACTCTTTCCCTACACGACGCTCTTCCGATCTTCTCAGTTTTTTTTTTTTTTTTTTTTTTTTTTTTTT(SEQIDNO:24)
ACACTCTTTCCCTACACGACGCTCTTCCGATCTTCAGACTTTTTTTTTTTTTTTTTTTTTTTTTTTTTT(SEQIDNO:25)
ACACTCTTTCCCTACACGACGCTCTTCCGATCTGACTCATTTTTTTTTTTTTTTTTTTTTTTTTTTTTT(SEQIDNO:26)
ACACTCTTTCCCTACACGACGCTCTTCCGATCTCACAGTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTT(SEQIDNO:27)
ACACTCTTTCCCTACACGACGCTCTTCCGATCTACACTGTTTTTTTTTTTTTTTTTTTTTTTTTTTTTT(SEQIDNO:28)
ACACTCTTTCCCTACACGACGCTCTTCCGATCTGCAGTATTTTTTTTTTTTTTTTTTTTTTTTTTTTTT(SEQIDNO:29)
ACACTCTTTCCCTACACGACGCTCTTCCGATCTGCTGATTTTTTTTTTTTTTTTTTTTTTTTTTTTTTT(SEQIDNO:30)
ACACTCTTTCCCTACACGACGCTCTTCCGATCTTAGTCGTTTTTTTTTTTTTTTTTTTTTTTTTTTTTT(SEQIDNO:31)
ACACTCTTTCCCTACACGACGCTCTTCCGATCTATGACGTTTTTTTTTTTTTTTTTTTTTTTTTTTTTT(SEQIDNO:32)
ACACTCTTTCCCTACACGACGCTCTTCCGATCTCGATGTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTT(SEQIDNO:33)
ACACTCTTTCCCTACACGACGCTCTTCCGATCTAGATCGTTTTTTTTTTTTTTTTTTTTTTTTTTTTTT(SEQIDNO:34)
ACACTCTTTCCCTACACGACGCTCTTCCGATCTTACGAGTTTTTTTTTTTTTTTTTTTTTTTTTTTTTT(SEQIDNO:35)
ACACTCTTTCCCTACACGACGCTCTTCCGATCTCGTAGATTTTTTTTTTTTTTTTTTTTTTTTTTTTTT(SEQIDNO:36)
ACACTCTTTCCCTACACGACGCTCTTCCGATCTTGTACGTTTTTTTTTTTTTTTTTTTTTTTTTTTTTT(SEQIDNO:37)
ACACTCTTTCCCTACACGACGCTCTTCCGATCTATCGTGTTTTTTTTTTTTTTTTTTTTTTTTTTTTTT(SEQIDNO:38)
ACACTCTTTCCCTACACGACGCTCTTCCGATCTGTGCATTTTTTTTTTTTTTTTTTTTTTTTTTTTTTT(SEQIDNO:39)
ACACTCTTTCCCTACACGACGCTCTTCCGATCTGAGCTATTTTTTTTTTTTTTTTTTTTTTTTTTTTTT(SEQIDNO:40)
ACACTCTTTCCCTACACGACGCTCTTCCGATCTCTCACAATTTTTTTTTTTTTTTTTTTTTTTTTTTTTT(SEQIDNO:41)
ACACTCTTTCCCTACACGACGCTCTTCCGATCTTACTGCATTTTTTTTTTTTTTTTTTTTTTTTTTTTTT(SEQIDNO:42)
ACACTCTTTCCCTACACGACGCTCTTCCGATCTATCAGCATTTTTTTTTTTTTTTTTTTTTTTTTTTTTT(SEQIDNO:43)
ACACTCTTTCCCTACACGACGCTCTTCCGATCTCGACTAATTTTTTTTTTTTTTTTTTTTTTTTTTTTTT(SEQIDNO:44)
ACACTCTTTCCCTACACGACGCTCTTCCGATCTCGTCAATTTTTTTTTTTTTTTTTTTTTTTTTTTTTTT(SEQIDNO:45)
ACACTCTTTCCCTACACGACGCTCTTCCGATCTCTCGAATTTTTTTTTTTTTTTTTTTTTTTTTTTTTTT(SEQIDNO:46)
ACACTCTTTCCCTACACGACGCTCTTCCGATCTTAGCACATTTTTTTTTTTTTTTTTTTTTTTTTTTTTT(SEQIDNO:47)
ACACTCTTTCCCTACACGACGCTCTTCCGATCTTCTAGCATTTTTTTTTTTTTTTTTTTTTTTTTTTTTT(SEQIDNO:48)
ACACTCTTTCCCTACACGACGCTCTTCCGATCTCACGTAATTTTTTTTTTTTTTTTTTTTTTTTTTTTTT(SEQIDNO:49)
ACACTCTTTCCCTACACGACGCTCTTCCGATCTATGCTCATTTTTTTTTTTTTTTTTTTTTTTTTTTTTT(SEQIDNO:50)
ACACTCTTTCCCTACACGACGCTCTTCCGATCTGCTACAATTTTTTTTTTTTTTTTTTTTTTTTTTTTTT(SEQIDNO:51)
ACACTCTTTCCCTACACGACGCTCTTCCGATCTGCATCTATTTTTTTTTTTTTTTTTTTTTTTTTTTTTT(SEQIDNO:52)
ACACTCTTTCCCTACACGACGCTCTTCCGATCTACATGCATTTTTTTTTTTTTTTTTTTTTTTTTTTTTT(SEQIDNO:53)
ACACTCTTTCCCTACACGACGCTCTTCCGATCTACGTCAATTTTTTTTTTTTTTTTTTTTTTTTTTTTTT(SEQIDNO:54)
ACACTCTTTCCCTACACGACGCTCTTCCGATCTCATGCTATTTTTTTTTTTTTTTTTTTTTTTTTTTTTT(SEQIDNO:55)
ACACTCTTTCCCTACACGACGCTCTTCCGATCTCTAGTCATTTTTTT...
【专利技术属性】
技术研发人员：欧阳宏伟，吴兵兵，李余，邹晓晖，
申请(专利权)人：浙江大学，
类型：发明
国别省市：浙江;33
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