RealHelix™ Direct qPCR Kit [Green] 은 DNA 정제 과정 없이 동물 조직, 식물 조직, 및 다양한 임상 시료 (혈액, 혈청, 소변, 머리카락 및 swab 시료 등)로부터 직접 real-time PCR을 수행할 수 있는 제품입니다. 본 제품의 2x Direct qPCR Premix [Green] 은 intercalating dye로 SYBR Green I이 적용되었으며, 항체 결합형 Taq 중합효소 (antibody-coupled hot-start Taq polymerase), dNTPs, MgCl₂, SYBR Green, 안정제(stabilizer)를 포함하고 있습니다. 특히, 본 제품은 시료에서 유래된 다양한 PCR 반응 억제 물질(PCR inhibitors)에 대한 내성을 나타내는 나노헬릭스만의 독특한 buffer system이 적용되어 직접 시료 적용과 함께 혈액과 같은 시료에 의한 형광검출 방해를 최소화하였습니다.
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.
