REPLI-g Advanced DNA Single Cell Kit(96)-汇佰在线平台

热搜推荐

推荐热卖

臻合智造手持式单道移液器（整支灭菌 )ZHP2.5

￥200.00

臻合智造手持式单道移液器（整支灭菌 )ZHP10

￥200.00

臻合智造手持式单道移液器（整支灭菌 )ZHP100

￥200.00

臻合智造手持式单道移液器（整支灭菌 )ZHP200

￥200.00

臻合智造手持式单道移液器（整支灭菌 )ZHP1000

￥200.00

舒适型乳胶手套.无粉.麻面.大号

￥41.88

舒适型乳胶手套.无粉.麻面.中号

￥41.88

舒适型乳胶手套.无粉.麻面.小号

￥41.88

MTS Cell Proliferation Assay K...

￥600.00

轻柔型丁腈手套.无粉.中号.蓝紫

￥36.25

商品名称：REPLI-g Advanced DNA Single Cell Kit(96)

货号：150365

[

Features

WGA with complete genome coverage from single eukaryotic cells or 1 ng genomic DNA
Simple 3-step protocol requires only 2 hours and 20 minutes, start to finish
Consistent yields of up to 25–35 µg DNA from 1 eukaryotic cell
Includes cryo-protect reagent to minimize DNA damage during single cell capture, storage or shipping

Product Details

DNA sequence analysis and genotyping of biological samples using next-generation sequencing (NGS) platforms is often limited by the amount of available sample. The REPLI-g Advanced DNA Single Cell Kit has been specifically optimized for use in sensitive microarray and NGS applications, providing high uniformity, with minimal allelic dropout, when starting with single cells or purified genomic DNA. Buffers and reagents undergo a unique, controlled decontamination procedure to minimize contaminating DNA and ensure highly reliable results. Accurate amplification of genomes with negligible sequence bias and minimal genomic dropouts is achieved with innovative Multiple Displacement Amplification (MDA) technology.

Performance

Genotyping and DNA sequence analysis of biological samples can be limited by a small amount of available sample. The REPLI-g Advanced DNA Single Cell Kit allows uniform amplification of whole genomic DNA from limited sample amounts and enables a greater variety and number of analyses to be performed. The average product length of amplified DNA is typically more than 10 kb, with a range between 2 kb and 100 kb, enabling all downstream applications such as complex genetic analysis, including long-range copy number variations, to be performed (1). DNA amplified with the REPLI-g Advanced DNA Single Cell Kit is highly suited for next-generation sequencing (NGS), array CGH genotyping applications or qPCR analysis.
Typical DNA yields from REPLI-g Advanced DNA Single Cell Kit amplifications are approximately 25–35 µg per 50 µl reaction. Depending on the quality of the input cell and its DNA, the resulting amount of amplified DNA may be less (fragmented or damaged gDNA should not be used). For best amplification results, we recommend collecting or storing cells in REPLI-g Advanced sc Storage Buffer (included with kit or available separately).

Principle

The REPLI-g Advanced DNA Single Cell Kit contains an optimized Phi 29 polymerase formulation, as well as buffers and reagents, for whole genome amplification (WGA) from single eukaryotic cells, very small amounts of sample or purified genomic DNA using Multiple Displacement Amplification (MDA). Phi 29 polymerase, a phage-derived enzyme, is a DNA polymerase with 3'→5' exonuclease activity (proofreading activity) that delivers up to 1000-fold higher fidelity compared to Taq DNA polymerase. Supported by the unique, optimized REPLI-g Advanced DNA Single Cell buffer system, Phi 29 polymerase easily solves secondary structures such as hairpin loops, thereby preventing slipping, stoppage and dissociation of the polymerase during amplification. The REPLI-g Advanced DNA Single Cell Kit is an improved version of the REPLI-g Single Cell Kit and includes an optimized reaction chemistry, a new single cell storage buffer and an improved protocol to increase the uniformity of amplification and reduce potential amplification bias. In addition, the MDA reaction time has been shortened to 2 hours.

Procedure

The REPLI-g Advanced DNA Single Cell Kit provides highly uniform amplification across the entire genome, with negligible sequence bias. The method is based on Multiple Displacement Amplification (MDA) technology, which carries out isothermal genome amplification utilizing a uniquely processive DNA polymerase capable of replicating up to 100 kb without dissociating from the genomic DNA template. In contrast to PCR-based methods, Phi 29 polymerase has a 3'–5' exonuclease proofreading activity to maintain 1000-fold higher fidelity than Taq Polymerase during replication. MDA technology is used in the presence of exonuclease-resistant primers to achieve high yields of DNA product from all kinds of eukaryotic tissues.
Genetic analyses often require substantial amounts of genomic DNA. Whole genome amplification solves the problem of low DNA quantity when starting with 1–1000 cells. The REPLI-g Advanced DNA Single Cell Kit overcomes these limitations by using a simple and reliable method to achieve accurate genome amplification. In addition, all components provided with the REPLI-g Advanced DNA Single Cell kit are exposed to UV radiation in a standardized procedure to minimize contaminating DNA. Reaction setup takes only 20 minutes and 3 steps, making the REPLI-g Advanced DNA Single Cell Kit protocol easy and reliable.
In the first step of the procedure, the cell sample is lysed under isothermal alkaline conditions and the cell is denatured. Cell lysis at room temperature reduces the number of possible DNA breaks and therefore improves uniformity of whole genome amplification. After denaturation has been stopped by the addition of neutralization buffer, a master mix containing buffer and DNA polymerase is added. The isothermal amplification reaction proceeds for 2 hours at 30°C. Because of the shortened MDA reaction time, the REPLI-g Advanced DNA Single Cell Kit enables single cell collection, WGA and even downstream analysis to be performed in a single day.

Applications

Downstream applications and instrumentation
Application	Instrumentation
Whole genome sequencing	Next-generation sequencing platforms*
Exome sequencing	Next-generation sequencing platforms*
SNP genotyping arrays	Array platforms*
Array CGH	Array platforms*
qPCR/PCR technologies	Real-time PCR/PCR cyclers*
Sanger sequencing	Capillary sequencers*
Pyrosequencing	PyroMark (QIAGEN)

* Available from various suppliers.

Learn more about our single cell products by visiting our Single Cell Resource.

References

1. Hosono, S. et al. (2003) Unbiased whole-genome amplification directly from clinical samples. Genome Res. 13, 954.

]

商品评价

100%
好评（100%）

中评（0%）

差评（0%）
推荐点：

买家评论事项：购买后有什么问题，满意，或者不不满，都可以在这里评论出来，这里评论全部源于真实的评论。
您可以对已购的商品进行评价
发表评论
详见积分中心积分规则

售后保障

退款说明

收到货物后如出现质量问题，买方需在收到货物7个工作日内向卖方提出书面异议。买方逾期不提出异议，视为货物符合其要求，卖方不付任何责任
出现质量以外的问题，如破损等，买方应在收货时提出，买方未提出异议，视为该产品合格。
特殊条款：如有以下情况造成延期交货，卖方不负任何责任：
如遇国内外战争。政治因素、自然条件等不可抗力因素造成的;
如国内外重大节日、国家法定节假日导致顺延的;
生产厂家自身原因造成的;
对于产品质量问题的解决仅限于产品本身，对其间接的影响，本公司不予承担。

全部咨询商品咨询支付配送售后

温馨提示：因产线可能更改商品包装、产地、附配件等未及时通知，且每位咨询者购买、提问时间等不同。为此，客服给到的回复仅对提问者3天内有效，其他网友仅供参考！给您带来的不便还请谅解，谢谢！

发表咨询

商品咨询：商品咨询支付配送售后

姓名:

电话:

邮箱:

地址:

研究方向：其它

验证码:

试剂

耗材

通用仪器

高端仪器

服务

REPLI-g Advanced DNA Single Cell Kit(96)

看了又看换一换

热搜推荐

推荐热卖

Features

Product Details

Performance

Principle

Procedure

Applications

属性

商品评价

售后保障

退款说明

问大家

联系我们