HelixAmp™ Direct PCR [3G]는 혈액, 동∙식물등 조직 시료로부터 DNA 정제 과정을 거치지 않고 target DNA를 증폭할 수 있는 제품입니다. 본 제품은 Taq polymerase, dNTP, 및 MgCl₂와 조직시료로부터 나오는 다양한 PCR 억제 물질(inhibtitor)에 내성을 갖는 Buffer가 2x 농도로 혼합된 premix 제품입니다. Pfu polymerase 계열 효소가 적용된 다른 제품과 달리, 3’→5’ exonuclease 기능이 없는 Taq polymerase가 적용된 본 제품은 다양한 유전자형 분석(Genotyping analysis)과 allele-specific PCR 분석이 가능합니다. 또한, Uracil-DNA-glycosylase(UDG) 적용이 가능하여 PCR 반응에서 빈번히 발생되는 carryover contamination을 방지할 수 있고 본 제품은 UDG/dUTP가 적용된 제품과 적용되지 않은 제품으로 구성되어 실험 목적에 따라 선택 하여 사용할 수 있습니다.
Figure 1. Direct PCR from various mouse tissues. Lysates directly extracted from each tissue without any DNA purification were used as templates.
1: Heart, 2: Liver, 3: Large intestine, 4: Small intestine, 5: Kidney, 6: Stomach, 7: Ear, 8: Brain, 9: spleen, NTC: No template control.
Figure 2. Direct PCR from whole blood and saliva. Human whole blood or saliva was used as the template for direct PCR without any pre-treatment.
Lane 1~10 : Amount of whole blood or saliva as the template (μl),
PTC: Purified genomic DNA, 10 ng was used as the template, NTC: No template control.
Figure 3. Direct PCR from various plant tissues. Lysates were directly extracted from each plant tissue without any DNA purification and used as the templates.
Direct PCRs were performed with a primer set specific for the chloroplast genome.
NTC : No template control.
Figure 4. Direct genotyping for selection of knock-out mice.
WT: Wild type mouse(339 bp), HZ: Heterozygotic mouse, KO: Knock-out mouse(589 bp), NTC: No template control.
Figure 5. Direct PCR for selection of transgenic plants. Lysates were directly extracted from each plant leaf and used as the template for PCR detection of transgenic plants.
Figure 6. Direct PCR and RFLP analysis for Zebrafish genotyping. Target DNAs were amplified directly from zerafish’s fin without DNA purification.
RFLP analysis was done using a restriction enzyme (Rsa I).
Application
- Allele-specific PCR
- PCR for genotyping
- PCR for selection of genetically modified organisms (GMO)
- Direct PCR amplification of target DNA without any DNA purification from various sample types such as whole blood, saliva, mouse tissues (tail, heart, liver, large intestine, small intestine, kidney, stomach, ear, brain, spleen), zebrafish’s fin, pork, beef, plant tissues (leaf and seed).
Contents
- 2x Direct PCR premix (with UDG or without UDG)
- 10x Dilution Buffer
- 6x Loading Dye
Quality control
HelixAmp™ Direct PCR [3G] is evaluated by amplification of PCR product corresponding to each sample type-specific DNA region directly using whole blood, leaf tissue, or animal tissue according to protocol.
