REPLI-g UltraFast Mini Kit

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。

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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	221,00 €	Add to cart
REPLI-g UltraFast Mini Kit (100) DNA Polymerase, Buffers, and Reagents for 100 x 20 μl ultrafast whole genome amplification reactions		150035	648,00 €	Add to cart

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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.

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

Application Notes

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	Genome-pULSe: Whole genome amplification & labeling for arrayCGH analysis - (EN)	Show details
	Oragene and REPLI-g Whole Genome Amplifi cation - (EN)	Show details

Brochures & Guides

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	Analyzing Genetic Differences - (EN) Second edition — innovative tools	Show details

FAQs

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	Can I reduce the amount of starting material below 10 ng when using the REPLI-g UltraFast Mini Kit? FAQ ID -1325	View
	Can I reduce the reaction time to e.g., 30 minutes, when using the REPLI-g UltraFast Mini Kit? FAQ ID -1326	View
	Why is there DNA in the no-template control reaction when using the standard REPLI-g procedure, but not when using the UltraFast procedure? FAQ ID -1327	View
	Will the REPLI-g UltraFast Mini Kit reaction always yield 10 µg DNA? FAQ ID -1328	View
	Do you have a protocol for cleanup of REPLI-g amplified DNA? FAQ ID -1545	View
	Can cDNA prepared with the QuantiTect Whole Transcriptome Kit be reampliied using REPLI-g Kits? FAQ ID -1611	View
	Where can I find background information and literature on Whole Genome Amplification with REPLI-g Kits? FAQ ID -1690	View
	What are possible reasons for reduced DNA yields with REPLI-g Kits? FAQ ID -2148	View
	What is REPLI-g whole genome amplification? FAQ ID -654	View
	Will REPLI-g work at high temperatures? FAQ ID -656	View
	What are the differences between MDA and DOP/PEP methods of Whole Genome Amplification? FAQ ID -665	View
	Does REPLI-g technology for whole genome amplification work with paraffin embedded samples? FAQ ID -673	View
	Why do I get amplification in a negative control DNA tube using the REPLI-g Kit for WGA? FAQ ID -675	View
	Is 2 kb the minimal gDNA fragment size for REPLI-g Whole Genome Amplification? FAQ ID -682	View
	What is the length of the MDA whole genome amplification product? FAQ ID -690	View
	What is the stability of the REPLI-g MDA product? FAQ ID -693	View
	How can I quantify the amount of REPLI-g DNA I have amplified? FAQ ID -694	View
	How can I determine the quality of my REPLI-g amplified products? FAQ ID -695	View
	Can I use REPLI-g for SNP Genotyping? FAQ ID -700	View
	Has anyone verified whole genome amplification accuracy with Sequencing? FAQ ID -701	View
	Are Centromeres and Telomeres amplified using REPLI-g WGA? FAQ ID -702	View
	What is the enzyme used in the REPLI-g reaction? FAQ ID -704	View
	Any data on the fidelity of the REPLI-g MDA technique? FAQ ID -707	View
	Can I purchase Phi29 DNA polymerase only? FAQ ID -708	View
	What are exo-resistant random hexamers used in the REPLI-g reaction? FAQ ID -710	View
	Can I use my own primers for REPLI-g WGA to amplify a specific chromosomal region? FAQ ID -712	View
	Will the random hexamers in the REPLI-g reaction interfere with downstream analysis? FAQ ID -713	View
	Do you have data on the DNA yield at different incubation time for the Repli-G Kits? FAQ ID -3113	View

Kit Handbooks

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	(EN) - REPLI-g UltraFast Mini Handbook For fast whole genome amplification from purified genomic DNA, blood, and cells	Show details

Scientific Posters

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	A novel approach to whole genome amplification and labeling of DNA samples for copy number variation detection on BAC microarrays	Show details
	Comparison of genotyping consistency between genomic and whole-genome amplified DNA using the Illumina GoldenGate and Infinium-II assays	Show details
	SNP genotyping of saliva DNA using Affymetrix GeneChip targeted genotyping system	Show details
	(EN) - The impact of whole genome amplification on forensic testing	Show details

Supplementary Protocols

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	Purification of REPLI-g amplified DNA using the QIAamp® DNA Mini Kit - (EN)	Show details
	Purification of DNA amplified using REPLI-g® Kits - (EN)	Show details

Safety Data Sheets

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	Safety Data Sheets Download Safety Data Sheets for QIAGEN product components.	View

Safety Data Sheets

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	MSDS REPLI-g UltraFast Mini Kit (25)
	MSDS REPLI-g UltraFast Mini Kit (100)

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