HelixAmp™ Direct PCR [3G] is designed for the PCR amplification directly from animal tissues, whole blood, and plant tissues without any DNA purification processes. This kit contains a 2x reaction mix including Taq DNA polymerase, dNTPs, MgCl₂, and unique buffer system to resist various PCR inhibitors of tissue samples. HelixAmp™ Direct PCR [3G] (Hot-start) is including hot-start Taq DNA polymerase that is inactive at lower temperature by an inhibitory antibody and using this enzyme avoids the polymerization from non-specifically bound primers during the setting of PCR mix and at the start of PCR cycles. At high temperature of the initial denaturation step of PCR, the inhibitory antibody is released by denaturation and the free Taq DNA polymerase becomes active. NanoHelix’s Direct PCR [3G] is based on Taq polymerase. Due to this enzyme’s robust amplification and 3’ to 5’ exo-negative properties, this kit could be used for allele specific PCR which is routinely used for various genotyping. Moreover the Uracil-DNA glycosylase with dUTP system for the prevention of carryover contamination can be applied to this Taq polymerase-based kit. dUTP could not be used with the Pfu DNA polymerase and its derivatives. A Uracil-DNA glycosylase and dUTP applied version is also available.

Figure 1. Direct PCR from various mouse tissues. Lysates directly extracted from each tissue without any DNA purification were used as template.

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 were use as template for direct PCR without any pre-treatment.

Lane 1~10 : Amount of whole blood or saliva as template(μl),

PTC: Purified genomic DNA, 10 ng was used as template, NTC: No template control.

Figure 3. Direct PCR from various plant tissues. Lysate directly extracted from each Plant tissues without any DNA purification were used as templates.

Direct PCRs were performed with a primer set specific for Chloroplast.

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 directly extracted from each plant leaves were used as template for PCR detection of transgenic plants.

Figure 6. Direct PCR and RFLP analysis for Zebrafish genotyping. Target DNA were amplified directly from zerafish’s fin without DNA purification.

RFLP analysis were 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.