HelixAmp™ Direct WGA Kit is a complete system for whole genome amplification from various tissues or samples directly without DNA purification processes. Very little amount of samples, several miligram or microliter volume, are required for the direct WGA. About 10 μg DNA products could be obtained in a standard reaction. The enzyme mix and buffer system are designed to tolerate against most amplification inhibitors found in crude samples. Phi29 DNA polymerase, the major polymerization enzyme of this kit, isothermally amplifies the genomic DNAs included in the samples with multiple displacement mechanism. Phi29 DNA polymerase could produce DNA strand up to 70 kb long with high fidelity. All required components including enzymes, buffers, dNTPs, random primers, and sample pretreatment reagents are supplied in this kit. The amplified DNA products could be applied for successive PCR, genotyping, and library construction. 

Figure 1. Direct WGA from various mouse tissues. (A). WGA products were amplified from very small(< 1 mg) piece of each tissue. (B). PCR detection of target DNA from 1 μl of serially diluted WGA products using the primer set for GAPDH.

Figure 2. Direct WGA from various plant seeds. (A). Direct WGA from various plant seeds without DNA purification. (B). PCR detection of target DNA from 1 μl of serially diluted WGA products using the primer set for a chloroplast-specific gene. 

Figure 3. Direct WGA from various plant leaves. (A). Direct WGA from various plant leaves without DNA purification. (B). PCR detection of target DNA from 1 μl of serially diluted WGA products using the primer set for RbcL. PTC: Control PCR using purified gDNA(10 ng/μl), NTC: from NTC product in direct WGA reaction. 

Figure 4. Direct WGA from Clinical and Forensic samples. (A). WGA products were amplified from various samples. (B). PCR detection of target DNA from 1 μl of serially diluted WGA products using the primer set for beta-actin or beta-globin. PTC: Control PCR using purified gDNA (10 ng/μl), NTC1: No template control, NTC2: from NTC product in WGA reaction. 

Application 

• Genotype analysis
• PCR and real-time PCR
• Construction of genomic library 

Quality control

Functional Assay : HelixAmp™ Direct WGA Kit was tested for the direct amplification from 1 μl of blood sample. 

Evaluation of amplified DNA 

• Agarose gel electrophoresis : The integrity of the amplified DNA is assessed by agarose gel electrophoresis.
• Quantification of amplified DNA : The DNA amplified with HelixAmp™ Direct WGA Kit are quantified using Quant-iT™ Assay Kit and Qubit fluorometer.