HelixAmp™ Genome Amplifier is a whole genome amplification kit suitable to amplify the whole genome from very little amount of genomic DNA sample. The whole genome amplification using HelixAmp™ Genome Amplifier is performed by Phi29 DNA polymerase with an isothermal MDA (multiple displacement amplification) based mode. Because of the strand displacement activity, strong 3’→5’ exonuclease activity and extreme processivity, Phi29 DNA polymerase could produce DNA up to 100 kb long with high fidelity. About 1,000 fold amplified DNA can be obtained from 10 ng genomic DNA within 90 min using this kit. The amplified genome can be directly applied, without further purification, to the downstream genetic analysis works including PCR, genotyping, and library constrictions.
Figure 1. WGA reactions from 10 ng of human genomic DNA using HelixAmp™ Genome Amplifier. Reactions were done in 20 ㎕ volume at 30℃ for 1.5 hour. 3 ㎕ of each reaction solutions were 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 reactions were determined with reaction time. About 8 ul 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 used 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 regions of chromosome 7, 8, 10, and 17 using the amplified whole genome as template. The amounts of template used in the 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 purified human genomic DNA as 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 test for the amplification of human whole genome using the 10 ng human genomic DNA at 30℃ for 1.5 hours