CN1221797A - 多聚酶链反应检测试剂盒Taq酶固相载体单人份检测管 - Google Patents

多聚酶链反应检测试剂盒Taq酶固相载体单人份检测管 Download PDF

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CN1221797A
CN1221797A CN 97109355 CN97109355A CN1221797A CN 1221797 A CN1221797 A CN 1221797A CN 97109355 CN97109355 CN 97109355 CN 97109355 A CN97109355 A CN 97109355A CN 1221797 A CN1221797 A CN 1221797A
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taq
taq enzyme
detector tube
solid phase
carrier
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CN1100150C (zh
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曾真
吴旗
詹群珊
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Abstract

一种多聚酶链反应检测试剂盒Taq酶固相载体单人份检测管及生产方法,是在单人份检测管中,采用固相载体,使高浓度的Taq酶按一定量附着其上,加入隔离相将其与扩增反应液水相分开,使Taq酶在检测实验前保持原始未稀释的状态,使试剂盒中Taq酶的稳定性达到与未实行单人份检测管技术前的技术指标,试剂盒稳定性和保存期都得到改善。

Description

多聚酶链反应检测试剂盒Taq酶固相载体单人份检测管
本发明涉及一种检测试剂盒及生产技术，具体地是一种多聚酶链反应检测试剂盒Taq酶固相载体单人份检测管及生产方法。
PCR技术的发明使当今世界分子生物技术的发展日新月异，令世人瞩目，在实际应用中为使操作更方便、实用，将检测试剂分装成单人份检测管已成发展趋势和定局，但现在单人份检测管仍使用Taq酶液相分装，这种方法对生产技术人员要求极高，费时费力，而且检测管间差异大，由于Taq酶被稀释成较低浓度，使试剂不易长期保存，而且Taq酶消耗大。
本发明目的旨在克服现有单人份检测管生产技术存在的不足，提供一种使试剂盒质量达到与未实行单人份检测管技术前的技术指标，使试剂盒稳定性和保存期都得到改善的多聚酶链反应检测试剂盒Taq酶固相载体单人份检测管生产方法。
本发明的目的是以下述方式实现的：一种多聚酶链反应检测试剂盒Taq酶固相载体单人份检测管，由附着Taq酶的固相载体将固相载体与扩增反应液分开的隔离相和扩增反应液组成。所述的固相载体为：对Taq酶无抑制作用；对Taq酶的吸附作用不会影响扩增反应；对扩增物无降解、水解作用；颗粒较均匀的固相物质，如玻璃珠、瓷珠等；所述的隔离相为：对Taq酶无抑制作用；不影响扩增反应；对扩增反应液的各种成份及产物无降解、水解作用的物质，如固态、半固态或液态矿物油、琼脂糖等。其生产方法是使Taq酶着在固相载体上。
本发明用颗粒均匀的小玻璃珠等为固相载体，使附着的Taq酶量较一致，因而检测管间差异小，因为固相载体上附着的为高浓度酶，且加入了隔离相将其与扩增反应液分开，使Taq酶在检测试验前保持原始的未稀释状态(不易失活)，易长期保存，减少Taq酶的消耗，使试剂盒的质量达到与未实行单人份检测管技术前的技术指标，且生产方法简单易学。
实施例一
1、将直径1mm的玻璃珠用1％盐酸浸泡过夜，用清水洗净、干燥后备用；
2、将处理过的玻璃珠按每个珠0.2μTaq酶的比例与5μ/μl Taq酶充分混合；
3、每个检测管(0.2ml EP管)中依次加吸附了Taq酶的玻璃珠1～2粒于管底，加入固体矿物油20μl，凝固后再加扩增反应液(含Tris-HCL缓冲液、mg+、dNTP、primer等)20μl。在其上再加20μl液态矿物油，该单人份检测管底层为着Taq酶的固相载体，第二层为隔离相，第三层为扩增反应液，第四层为液态隔离层。
实施例二
1、将直径1.5mm的瓷珠用1％盐酸浸泡16小时，用清水洗净、干燥备用；
2、将备用的瓷珠按每个珠0.2μTaq酶的比例与5μ/μl Taq酶充分混合；
3、每个检测管(0.5ml EP管)中依次加入25μl扩增反应液、25μl半固体矿物油和1～2粒吸附了Taq酶的瓷珠，该检测管底层为扩增反应液，第二层为隔离层，第三层为固相载体。

Claims (2)

1、一种多聚酶链反应检测试剂盒Taq酶固相载体单人份检测管，其特征在于所述检测管由吸附着Taq酶的固相载体，将固相载体与扩增反应液分开的隔离相和扩增反应液组成，所述的固相载体为：对Taq酶无抑制作用；对Taq酶的吸附作用不会影响扩增反应；对扩增物无降解、水解作用；颗粒较均匀的固相物质；所述的隔离相为：对Taq酶无抑制作用；不影响扩增反应；对扩增反应液的各种成份及产物无降解、水解作用的物质。
2、生产如权利要求1所述的Taq酶固相载体单人份检测管生产方法，其特征在于使Taq酶附着在固相载体上，放入单人份检测管中，再加入隔离相、扩增反应液，或在单人检测管中先加入扩增反应液，再依次加入隔离相和使Taq酶吸附其上的固相载体。
CN97109355A 1997-12-30 1997-12-30 多聚酶链反应检测试剂盒Taq酶固相载体单人份检测管 Expired - Fee Related CN1100150C (zh)

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Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN100429131C (zh) * 2003-09-17 2008-10-29 中山大学达安基因股份有限公司 设有内部分隔的试剂盒组分容器

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN100429131C (zh) * 2003-09-17 2008-10-29 中山大学达安基因股份有限公司 设有内部分隔的试剂盒组分容器

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