用于从珍贵或少量样本中超快速,高度均一地进行全基因组扩增

  • 快速全基因组扩增,仅需1–1.5小时
  • 获得无限量DNA适合多种下游应用
  • 从各种起始样本中进行高重复性扩增
  • 高度稳定的DNA适合长期储存

基因组DNA量不足以进行基因组分析,这一问题可通过样本内全基因组DNA扩增解决。REPLI-g UltraFast Kit包含REPLI-g UltraFast DNA Polymerase,该聚合酶可在60–90分钟内高度均一、正确的对少量样本进行全基因组扩增,通常20 μl反应体系产量可达7 μg。起始样本可以是基因组DNA、新鲜或干血、口腔拭子、新鲜或冷冻组织和培养细胞等。扩增得到的DNA可即用于多种基因分析。

Product Product no. Cat. no. List price:
 
 
Show details
    varies
Can't order online?
To place an order via phone, email or for requesting a quote, please provide the product’s name, number and catalog number.
Product Cat. no. List price:
REPLI-g UltraFast Mini Kit (25)
DNA Polymerase, Buffers, and Reagents for 25 x 20 μl ultrafast whole genome amplification reactions
150033
Inquire
REPLI-g UltraFast Mini Kit (100)
DNA Polymerase, Buffers, and Reagents for 100 x 20 μl ultrafast whole genome amplification reactions
150035
Inquire
The REPLI-g UltraFast Mini Kit is intended for molecular biology applications. This product is not intended for the diagnosis, prevention, or treatment of a disease.
  • Main Image Navi
Purified genomic DNA procedure.|Consistent and high yields of amplified DNA.|
Amplification of genomic DNA using the REPLI-g UltraFast Mini Kit involves 3 basic steps. First, the sample undergoes gentle alkaline denaturation. Next, the sample is neutralized, and finally incubated with REPLI-g UltraFast DNA Polymerase at 30°C.|Genomic DNA samples (10 ng each) and non-template control (NTC) samples (containing no template DNA) were amplified using the REPLI-g UltraFast Mini Kit and the DNA yield was determined after various times. An average yield of 7 µg was obtained after amplification for 60 minutes, while 10–13 µg DNA was obtained after 90 minutes. Up to 5 µg DNA was obtained after just 45 minutes, allowing even faster downstream genetic analysis. No DNA was detected in the non-template control reactions.|
Performance
孵育90分钟后,可从20 µl反应体系中获得7 µl DNA。如果时间有限,45分钟后也可获得足以用于下游应用的产物(参见"Consistent and high yields of amplified DNA")。阴性对照样本(无模板DNA)不被扩增,能够可靠检测样本或污染DNA是否存在(参见"Consistent and high yields of amplified DNA")。
Principle

用于基因组分析的基因组DNA量通常十分有限。全基因组扩增技术通过对样本中的所有DNA进行全基因组扩增,为下游应用提供足量的DNA,克服这一问题。REPLI-g UltraFast技术可高度均一的进行全基因组扩增,序列偏差小。该方法基于MDA技术,采用独特的DNA聚合酶进行等温基因组扩增,可在较短时间内扩增得到长达100 kb的DNA产物,避免了基于PCR的扩增方法中常见的高序列偏差。此外,独特的碱性变性缓冲液确保了基因组DNA的温和变性,避免热变性导致的模板DNA片段化。因此可确保在下游应用中获得更加灵敏、更加可靠的结果。

Procedure
使用REPLI-g UltraFast Mini Kit进行全基因组扩增需要3个基本步骤。首先,对样本(10 ng纯化的基因组DNA、0.5 µl全血或300个组织培养细胞)进行温和的碱变性,避免模板DNA片段化。然后中和样本,加入REPLI-g UltraFast DNA Polymerase于30°C孵育。60分钟后可获得产物,扩增得到的DNA无需纯化,可即用于下游应用。
Applications

REPLI-g扩增得到的基因组DNA适合多种下游应用,包括:

  • 基因分型(如:SNP、STR、微阵列)
  • 终点法PCR、real-time PCR
  • 测序 
  • 比较基因组杂交 
Feature
Specifications
Amplification Whole genomic DNA
Applications Genotyping, microarray, PCR, real-time PCR
Denaturation step Alkaline
Maximum input volume 10 ng DNA, 0.5 µl whole blood, ~300 cells/µl
Minimal pipetting volume needed 1 µl
Quality assessment No
Reaction time 60–90 minutes
Reaction volume 20 µl
Samples per run; throughput Mid
Starting amount of DNA >10 ng purified genomic DNA
Starting material Genomic DNA, blood, cells
Technology Multiple Displacement Amplification (MDA)
Yield ~10 µg

You are not authorized to download the resource

Brochures & Guides
1
FAQs
28
FAQ ID -1325
View
FAQ ID -1326
View
FAQ ID -1327
View
FAQ ID -1328
View
FAQ ID -1545
View
FAQ ID -1611
View
FAQ ID -1690
View
FAQ ID -2148
View
FAQ ID -3113
View
FAQ ID -654
View
FAQ ID -656
View
FAQ ID -665
View
FAQ ID -673
View
FAQ ID -675
View
FAQ ID -682
View
FAQ ID -690
View
FAQ ID -693
View
FAQ ID -694
View
FAQ ID -695
View
FAQ ID -701
View
FAQ ID -702
View
FAQ ID -704
View
FAQ ID -707
View
FAQ ID -708
View
FAQ ID -710
View
FAQ ID -712
View
FAQ ID -713
View
FAQ ID -700
View
Kit Handbooks
1
For fast whole genome amplification from purified genomic DNA, blood, and cells
Show details