HelixAmp™ Genome Amplifier (WGA)는 Phi29 DNA polymerase를 이용하여 극소량의 genomic DNA(~10 ng)로부터 1.5 ~ 2 시간 반응으로 1,000 배 이상 whole genome을 증폭할 수 있는 제품입니다. 본 제품에 사용된 Phi29 DNA polymerase는 MDA(multiple displacement amplification) 및 3’→5’ exonuclease 활성을 가지고 있어서 높은 정확도로 전체 genomic DNA를 70 ~ 100 kb 크기로 증폭할 수 있으며, 본 제품으로 증폭된 genomic DNA는 유전자형 분석(Genotyping analysis), 희귀유전자원의 확보 및 보존, 그리고 genomic DNA library 제작 등 다양한 DNA 분석 실험에 사용 될 수 있습니다.
Figure 1. WGA reactions from 10 ng of human genomic DNA using HelixAmp™Genome Amplifier. Reactions were performed in 20 ㎕ volume at 30℃ for 1.5 hours. 3 ㎕ of each reaction solution was loaded on a 1% agarose gel for analysis.
Figure 2. Time-course measurements of DNA amplification using HelixAmp™ Genome Amplifier from 10 ng of genomic DNA purified from blood collections. Ten ng of Genomic DNA samples were amplified using the HelixAmp™Genome Amplifier, and the DNA yields of each reaction were determined by reaction time. About 8 ul of products were obtained in 60 minutes and 12 ul after 90 min incubations.
Figure 3. Validation of amplified genome products by chromosome-targeting PCR analysis. Ten ng of human genomic DNA, isolated from blood collection, was applied in the whole genome amplification using HelixAmp™Genome Amplifier. (A) PCR detections of human beta-actin gene from the amplified genomes. Purification is not required for the PCRs using the products of HelixAmp™Genome Amplifier.(B) PCR detections of each chromosome-specific region of chromosome 7, 8, 10, and 17 were performed using the amplified whole genome as the template. The amounts of the template used in each 50 ㎕ PCR reactions are: lane 1, 1 ㎕; lane 2; 0.1 ㎕, lane 3; 0.01 ㎕ of the amplified genome product and lane 4; 1 ng of the purified human genomic DNA as a control.
Application
- SNP or SSR genotype analysis
- PCR and real-time PCR
- Construction of genomic library
Contents
- Enzyme Mix with Phi29 DNA polymerase:
- 8x DB
- 10x NB
- dNTP Mix (each 10 mM)
- Primer Mix
- Reaction Buffer
Quality Control
HelixAmp™ Genome Amplifier was tested for the amplification using 10 ng human genomic DNA at 30℃ for 1.5 hours.
