CN109536591A - 一种pcr扩增方法 - Google Patents
一种pcr扩增方法 Download PDFInfo
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- 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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- 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
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- 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
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。
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Citations (3)
Publication number | Priority date | Publication date | Assignee | Title |
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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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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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