RealHelix™ 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™ Direct qPCR Kit [Green] from blood samples.
A lysate of whole blood was prepared following the protocol, and 1 μl of the lysate was used as a template in this reaction containing an intercalating dye, SYBR Green l. 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™ Direct qPCR Kit [Green] from lambda DNA-spiked serum samples.
A lysate from the lambda DNA-spiked serum was prepared following the protocol. 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™ Direct qPCR Kit [Green] from urine sample.
Cell pellets from 1 ml of urine by centrifugation and PBS-wash were used for the preparation of lysate following the protocol. 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™ Direct qPCR Kit [Green] from buccal swab sample.
The lysate was prepared from a tissue sample collected by a buccal swab following the protocol. 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 displayed. NTC, no template control. Each reaction was duplicated.
Figure 5. Real-time PCR using RealHelix™ Direct qPCR Kit [Green] from mouse tail sample.
1 mm of mouse tail was used for the preparation of lysate following the protocol. 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.
