计算溶液所需的质量、体积或浓度。
为什么选择此级别
，适用于对基线干扰要求严格的色谱和分析工作流程。
储存与运输
-20°C储存,避免反复冻融。超低温运输 。请查阅批次 COA 获取详细规格。
质量文档
SDS、COA、产品数据表及规格说明书均可下载。可通过批号查询获取批次 COA。
文献证明
在色谱分析、有机合成和交叉偶联反应领域已被 0 篇同行评审文献引用。
概述
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产品简介
单细胞全基因组扩增试剂盒可以以单个细胞或者微量样本为模板实现全基因组扩 增。单细胞扩增反应时间短，总过程约3 h，反应经裂解、预扩增、扩增过程即可得到 2-5 μg的基因组DNA，大小在200-1500 bp左右。扩增产物可广泛适用于二代测序、大 片段拷贝数变异分析、SNP分型、qPCR分析、基因芯片分析等。】
自备仪器、耗材
PCR仪
反应管：建议使用低吸附的管子
枪头：建议使用高质量过滤枪头
微型离心机、漩涡混和仪
注意事项
本产品检测灵敏度极高，实验操作应在正压的超净工作台或洁净环境中完成，扩增 反应产物浓度较高，应做好隔离，避免扩增产物导致的气溶胶污染。
操作流程示意图
操作步骤
实验前准备
通过流式细胞仪分选，缓冲液稀释，显微操作和激光显微切割等方式获得单细胞， 建议在实验前对细胞进行清洗，清洗液为不含Mg2+和Ca2+的1×PBS溶液，需注意，确 保后续实验中的PBS溶液的体积不超过2 µl。
注意
由于实验全程在同一PCR管中进行且反应体积较小，因此加液操作时移液器吸头不 要接触管中液体，避免将单细胞或DNA带出反应体系；移液时，请沿管壁小心添加，勿 吹打PCR管中液体；反应前，请进行短暂离心，确保反应体系中的液体混和均匀。 使用前请将细胞裂解酶、预扩增酶和扩增酶置于冰上解冻。
1．细胞裂解
1）根据反应的数量N，混和Cell Lysis Buffer和Cell Lysis Enzyme，震荡混匀，短 暂离心备用。
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2）将单细胞与细胞裂解混合液混匀于PCR管中，运行以下程序。
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2．预扩增反应
1）根据反应的数量N，混和Pre-Amp Buffer和Pre-Amp Enzyme Mix，震荡混匀，短 暂离心备用。
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2）加入5 μl预扩增混合液于上步10 μl裂解反应产物中，运行以下程序。
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3．扩增反应
1）根据反应的数量N，混和Amplification Buffer和Amp Enzyme Mix，震荡混匀，短 暂离心备用。
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2）加入60 μl扩增混合液于上步15 μl预扩增反应产物中，运行以下程序。
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说明：循环数可根据需要调整，对于流式分选等方式获得的单细胞建议14个循环。
扩增产物检测
1．琼脂糖凝胶电泳
取5 μl扩增产物进行琼脂糖凝胶电泳（1%琼脂糖凝胶，110 V，25-35分钟）， 扩增 产物大小为200-1500 bp。
2．定量
对扩增产物进行磁珠或柱式纯化，纯化产物使用Qubit进行定量，终产量为2-5 μg。
Products content
Products Introduction
The Single Cell Whole Genome Amplification Kit can be used as a template for whole genome amplification of single cells or micro samples. The total time for single-cell amplification is about 3 hours, and 2-5 μg of genomic DNA, with a size of 200-1500 bp, can be obtained after lysis, pre-amplification and amplification. The amplified product can be widely used in second-generation sequencing, large fragment copy number variation analysis, SNP typing, qPCR analysis and gene chip analysis.
Bring your own instruments and reagents
PCR instrument
Reaction tubes: low adsorption tubes recommended
Gun Heads: High quality filtered gun heads are recommended
Microcentrifuge, vortex mixer
caveat
The sensitivity of this product is very high, the experimental operation should be completed in a positive pressure ultra-clean bench or clean environment, the concentration of the amplification reaction products is high, should be well isolated to avoid aerosol contamination caused by amplification products.
Operation flow diagram
procedure
Pre-experiment preparation
Single cells were obtained by flow cytometry sorting, buffer dilution, micromanipulation and laser microdissection. It is recommended that the cells be washed prior to the experiments with a 1× PBS solution free of Mg2+ and Ca2+ , taking care to ensure that the volume of PBS solution in subsequent experiments does not exceed 2 µl.
take note of
Since the whole experiment is carried out in the same PCR tube and the reaction volume is small, the pipette tip should not touch the liquid in the tube when adding liquid, so as to avoid taking single cells or DNA out of the reaction system; when pipetting, please add the liquid along the wall of the tube carefully and do not blow the liquid in the PCR tube; before the reaction, please centrifuge briefly to make sure that the liquid in the reaction system is mixed evenly. Thaw the cell lysate, pre-amplifier and amplifier on ice before use.
cell lysis
1）Mix Cell Lysis Buffer and Cell Lysis Enzyme according to the number of reactions N, shake to mix, centrifuge briefly and set aside.
2）Mix single cells with the cell lysis mix in a PCR tube and run the following program.
2. Pre-amplification reaction
1）Mix Cell Lysis Buffer and Cell Lysis Enzyme according to the number of reactions N, shake to mix, centrifuge briefly and set aside.
2）Add 5 μl of pre-amplification mix to 10 μl of lysis reaction product from the previous step and run the following program.
3. Amplification reaction
1）Mix Amplification Buffer and Amp Enzyme Mix according to the number of reactions N, mix with shaking, centrifuge briefly and set aside.
2）Add 60 μl of amplification mix to 15 μl of pre-amplification reaction product from the previous step and run the following program.
Note: The number of cycles can be adjusted as needed, 14 cycles are recommended for single cells obtained by flow sorting, etc.
Amplification product detection
1. Agarose gel electrophoresis
5 μl of the amplified product was subjected to agarose gel electrophoresis (1% agarose gel, 110 V, 25-35 min), and the amplified product was 200-1500 bp in size.
2. Quantitative
Amplification products were subjected to magnetic bead or column purification, and purified products were quantified using Qubit with a final yield of 2-5 μg.
规格
别名
单细胞WGA试剂盒
英文名称
Single Cell WGA Kit
储存条件
-20°C储存,避免反复冻融
运输条件
超低温运输
产品线
生物产品
名称和识别符
分子类型
试剂盒
技术文档
高级数据
质检证书（CoA,COO,BSE/TSE 和分析图谱)
C of A & Other Certificates(BSE/TSE, COO):
输入批号以搜索分析图谱:
溶液计算器
产品仅提供给经过验证的企业、机构和合格的专业人员用于研发。不适用于人类、动物、诊断或治疗。
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