The RealHelix^TM Direct qPCR Kit [Green] is designed for an intercalating dye based qPCR amplification directly from animal tissues, plant tissues , and various clinical samples including whole blood, serum, urine, hair and swab collections without any DNA purification processes. The 2x Direct qPCR Premix [Green] in this kit contains antibody inhibited Taq DNA polymerase, dNTPs, MgCl₂ , SYBR Green, stabilizer, and unique buffer system to resist various PCR inhibitors of tissue samples.

Figure 1. Real-time PCR using RealHelix^TM Direct qPCR Kit [Green] from blood samples. A lysate of whole blood was prepared following the protocol of this kit and 1 μl of the lysate was used as template in this reaction containing an intercalating dye, SYBR Green l. Ten (10) ng of human genomic DNA was used for a control reaction on a Real-time PCR instrument, Bio-Rad CFX96. The amplification curves (upper panels) and melting analysis results (lower panels) are shown. NTC, no template control. Each reaction was duplicated.

Figure 2. Real-time PCR using RealHelix^TM Direct qPCR Kit [Green] from lambda DNA-spiked serum samples. A lysate from the lambda DNA-spiked serum was prepared following the protocol of this kit. 1 μl of prepared lysate(test sample) and 1 pg of lambda DNA(positive control) were analyzed with a lambda DNA-specific primer set and an intercalating dye, SYBR Green l on a Real-Time PCR instrument, Bio-Rad CFX96. The amplification curves (upper panels) and melting analysis results (lower panels) are shown. NTC, no template control. Each reaction was duplicated.  

Figure 3. Real-time PCR using RealHelix^TM Direct qPCR Kit [Green] from urine sample. Cell pellets from 1 ml urine by centrifugation and PBS-wash were used for the preparation of lysate following the protocol of this kit. 1 μl of prepared lysate(test sample) and 10 ng of human genomic DNA (positive control) were analyzed with a human beta-globin gene-specific primer set and an intercalating dye, SYBR Green l. Direct qPCR was done on a Real-Time PCR instrument, Bio-Rad CFX96. The amplification curves (upper panels) and melting analysis results (lower panels) are shown. NTC, no template control. Each reaction was duplicated.

Figure 4. Real-time PCR using RealHelix^TM Direct qPCR Kit [Green] from buccal swab sample. The lysate was prepared from a tissue sample collected by a buccal swab following the protocol of this kit. 1 μl of prepared lysate(test sample) and 10 ng of human genomic DNA (positive control) were analyzed with a human beta-globin gene-specific primer set and an intercalating dye, SYBR Green l on a Real-Time PCR instrument, Bio-Rad CFX96. The amplification curves (upper panels) and melting analysis results (lower panels) are shown. NTC, no template control. Each reaction was duplicated. 

Figure 5. Real-time PCR using RealHelix^TM Direct qPCR Kit [Green] from mouse tail sample. 1 mm size of mouse tail cut was used for the preparation of lysate following the protocol of this kit. 1 μl of prepared lysate(test sample) and 10 ng of mouse genomic DNA (positive control) were analyzed with a mouse sox21 gene-specific primer set and an intercalating dye, SYBR Green l on a Real-Time PCR instrument, Bio-Rad CFX96. The amplification curves (upper panels) and melting analysis results (lower panels) are shown. Each reaction was duplicated.  

Application 

• Direct and quantitative real-time PCR 

Contents 

• 2x Direct qPCR Premix [Green]
• 3x P-solution
• 10x Dilution Buffer