HelixAmp™ Direct WGA Kit는 극 소량의 조직 시료로부터 핵산의 분리 및 정제 과정없이 직접 ug/ul농도 수준으로 전체 genomic DNA를 증폭할 수 있는 제품으로, 동,식물의 조직 시료, 혈액, 혈청, 타액 등 임상관련 시료, 그리고 모근이나 혈흔과 같은 법의학적 시료 등 다양한 시료에 적용 가능합니다. 본 제품은 1 ul 혈액 (조직 시료의 경우, 1 ~ 3 mm 크기) 시료로부터 표준 반응을 통해 대략 10 µg 정도의DNA 산물을 얻을 수 있으므로, 특히 극소량의 희귀 시료에 대한 유전자원 확보에 유용하게 이용될 수 있습니다. Multiple displacement 기능과 3’→5’ exonuclease활성을 가지고 있는 Phi29 DNA polymerase는 매우 정확하게 70 ~ 100 kb길이로 DNA를 증폭할 수 있고, 증폭된 genomic DNA는 유전자형 분석(Genotyping analysis), 희귀유전자원의 확보 및 보존, 그리고 genomic DNA library 제작 등 다양한 DNA 분석 실험에 사용될 수 있습니다.
Figure 1. Direct WGA from various mouse tissues. (A). WGA products were amplified from very small (< 1 mg) samples 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.
