In the situation that there is little amount of total DNA available and it is inaccessible to more amount of DNA samples, we can use whole genome amplification (WGA) technology to increase the amount of DNA, allowing research to be able to continue. Whole genome amplification is a technology that enables non-selective amplification of genomic DNA. It allows researchers to dramatically increase the total amount of genomic DNA without sequence-specific preference. Genesky holds independent intellectual property of its self-invented Whole Genome Amplification kit. This kit utilizes a special polymerase to reliably, efficiently and non-selectively amplify genomic DNA by the mode of ‘ultra branch amplification’. The polymerase has strong template binding ability, and is able to continuously extend over 70 kb along the template. Also, this polymerase possess 3’ - 5’ exonuclease activity, with error rate as low as 5X10-6, which is about 100 times lower than Taq Pol. This ensures the high fidelity of amplification.
Technique Highlights
– With 10ng (or even 1ng) genomic DNA as starting material, >15ug (20ul reaction system) or >35ug (50ul reaction system) amplification product may be expected.
– The amplification product can be up to 100kb in length, with average length of >10kb (see figure II), thus the amplification products can be used for cloning, haplotype determination and long PCR experiments.
– When the amount of DNA template is high enough (gDNA > 10g or cell number > 600), the amplification efficiency at each locus tends to be similar, so that amplification coverage approaches 100%.
– The kit uses a high-fidelity DNA polymerase, which means substantially low probability of mutation introduction or STR replication slippage during the process of amplification. Thus, the amplification products can be used for downstream DNA sequence analysis experiments such as sequencing, SNP and STR genotyping etc.
Sample Requirement
Genomics DNA, fresh or dried whole blood, oral mucosa scraping liquid, hair root and cultured cell can all be used as starting material for this kit. The kit requires as little as 5 ng genomics DNA for amplification, but taking into account of the systematic bias of DNA quantification and inconsistency of DNA quality between samples, the recommend amount for genomic DNA is >20 ng. Sufficient amount and quality of genomic DNA reduces the chance of unbalanced amplification of allelic genes.
Service Workflow
1. Whole genome amplification2. Quality control by agarose gel electrophoresis
Deliverables
1. Experiment record2. Quality check report
