CN109536591A - 一种pcr扩增方法 - Google Patents

一种pcr扩增方法 Download PDF

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Publication number
CN109536591A
CN109536591A CN201811475873.2A CN201811475873A CN109536591A CN 109536591 A CN109536591 A CN 109536591A CN 201811475873 A CN201811475873 A CN 201811475873A CN 109536591 A CN109536591 A CN 109536591A
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sample
reaction
classified
data
reaction table
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侯明飞
肖聪
朱月艳
孙子奎
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SHANGHAI PERSONAL BIOTECHNOLOGY CO Ltd
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SHANGHAI PERSONAL BIOTECHNOLOGY CO Ltd
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    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
    • C12Q1/6844—Nucleic acid amplification reactions
    • C12Q1/686—Polymerase chain reaction [PCR]

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Abstract

本发明公开了一种PCR扩增方法，其特征在于，包括如下步骤：样品分类标记步骤：将扩增V区相同的样品进行分类；反应表制作步骤：根据对应的样品、样品来源及引物信息统一记录在反应表中；样品反应步骤：将分类好的样品配置MIX后开始上机反应；样品数据处理步骤：将不同样品的下机数据保留，对相关参数做宏。本发明的有益效果在于：通过合理的进行样品分类标记，将散乱的大批量样本进行有效组合后再上机进行扩增，降低了实验操作错误的风险和降低了后续数据整理的难度。

Description

一种PCR扩增方法
技术领域
本发明涉及一种生物实验检测领域，具体涉及一种PCR扩增方法。
背景技术
PCR是一种体外核酸扩增技术，理论上可以在1-2h之内将目标DNA序列从几个拷贝扩增到几百万倍。
当前，每次实验样品总是有很多份，每次实验如果针对每个样品单次PCR 扩增不仅需要消耗大量的人力物力，且因扩增实验较多，容易在实际操作当中对扩增结果的标记错误，后期不易针对不同的数据进行比对、归纳和查找。
当前，随着基因测序领域的发展，对于PCR扩增的速度和准确度及数据的易处理性都有了更高的要求。因此，一种更加易于操作和便于收集处理数据的PCR扩增方法变得尤为重要。
发明内容
为了克服现有技术所存在的上述缺陷，本发明的目的在于一种PCR的扩增方法，可以达到如下三个目的：1.提高样品扩增的效率；2.简化数据统计和相关实验资料的查询3.减少扩增时，因样品太多带来的错误。
为了实现本发明的目的，所采用的技术方案是：一种PCR扩增方法，包括如下步骤：
样品分类标记步骤：将扩增V区相同的样品进行分类；
反应表制作步骤：根据对应的样品、样品来源及引物信息统一记录在反应表中；
样品反应步骤：将分类好的样品配置MIX后开始上机反应；
样品数据处理步骤：将不同样品的下机数据保留，对相关参数做宏。
本发明的有益效果在于：
通过合理的进行样品分类标记，将散乱的大批量样本进行有效组合后再上机进行扩增，降低了实验操作错误的风险和降低了后续数据整理的难度。
附图说明
图1为扩增方法的宏表格示意图1；
图2为扩增方法的宏表格示意图2；
图3为扩增后的结果示意图3。
具体实施方式
下面结合实施例对本发明做进一步说明：
一般情况下的PCR扩增是根据单一订单。将实验记录做好；进行扩增具体参见图1；
而现有技术当中，数据不便于统一管理，操作人员仅对自己的操作比较清楚。
见表1
而本发明的方法是依照下列步骤，来进行PCR扩增：
样品分类标记步骤：将扩增V区相同或扩增条件一样的样品进行分类；
反应表制作步骤：根据对应的样品、样品来源及引物信息统一记录在反应表中；
样品反应步骤：将分类好的样品配置MIX后开始上机反应；
样品数据处理步骤：将不同样品的下机数据保留，对相关参数做宏。
为了进一步说明本发明的技术方案，对本发明的技术方案进行举例参考：
举例参考：
首先将扩增的数据按扩增V区进行分类：
见表2
订单 客户 来源 类型 样品数 反应数 扩增V区
SP180930456 客户1 略 略 23 23 v34
略 略 23 18SV4
SP180930445 客户2 略 略 18 18 v34
SP180930460 客户3 略 略 3 3 v34
客户3 略 略 3 ITS1
SP180930452 客户4 略 略 9 9 v34
客户4 略 略 9 ITS1
SP180930454 客户5 略 略 5 5 v34
SP180930442 客户6 略 略 4 4 v45
SP180930440 客户7 略 略 6 6 v34
SP180930453 客户8 略 略 22 22 v34
2.将客户的样品信息黏贴到宏表格内，见图1，该表的设计是跟据前期提供的检测数据而来的：
3.将数据粘贴后，点击反应表出现如表3和图2：
4.做好反应表后，可以将相应的订单放好；根据反应表，第一步：将对应订单的样品找好，放在96孔双面板上，第二步：将对应的引物找好；第三步：根据反应表的个数计算所需mix；
5.将实验操作台收拾干净，实验台上仅留样品，引物，和实验用的耗材；
6.根据摆好的顺序，使用96孔板。开始做反应。
7.反应结束后，电泳：也是使用该反应表做记录。胶图名称与反应表名称一致；
8.胶图一块点48个样品，正好对应反应表的A-D行；命名为：反应表-1 第二块胶命名为：反应表-2，见图3；
9电泳图跑好，割胶回收后；补充完表格信息胶回收位置。
10.因为标准统计，可以将每个人所做的信息汇总在一起，后期可以检索相关扩增信息。

Claims (1)

1.一种PCR扩增方法，其特征在于，包括如下步骤：
样品分类标记步骤：将扩增V区相同的样品进行分类；
反应表制作步骤：根据对应的样品、样品来源及引物信息统一记录在反应表中；
样品反应步骤：将分类好的样品配置MIX后开始上机反应；
样品数据处理步骤：将不同样品的下机数据保留，对相关参数做宏。
CN201811475873.2A 2018-12-04 2018-12-04 一种pcr扩增方法 Pending CN109536591A (zh)

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Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN101035620A (zh) * 2004-04-08 2007-09-12 生物马特里卡公司 用于生命科学的样品存储和样品管理的综合
WO2009115761A2 (fr) * 2008-03-11 2009-09-24 Imagene Procede industriel d'encapsulation de materiel biologique en vue d'une conservation a temperature ambiante
WO2015116978A1 (en) * 2014-01-31 2015-08-06 Carnegie Mellon University Device and method for clinical data sampling and specimen banking

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN101035620A (zh) * 2004-04-08 2007-09-12 生物马特里卡公司 用于生命科学的样品存储和样品管理的综合
WO2009115761A2 (fr) * 2008-03-11 2009-09-24 Imagene Procede industriel d'encapsulation de materiel biologique en vue d'une conservation a temperature ambiante
WO2015116978A1 (en) * 2014-01-31 2015-08-06 Carnegie Mellon University Device and method for clinical data sampling and specimen banking

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Application publication date: 20190329