RealHelix™ Direct qPCR Kit [Green]은 동물 조직, 식물 조직 및 다양한 임상 시료 (혈액, 혈청, 소변, 모근 및 swab 시료 등)로부터 별도의 DNA 정제 과정 없이 intercalating dye를 이용한 real-time PCR을 할 수 있도록 디자인 된 제품입니다. 본 제품의 2x Direct qPCR Premix [Green] 은 antibody-mediated hot-start Taq 중합효소, dNTPs, MgCl₂, green fluorescent dye, 안정제(stabilizer)를 포함하고 있습니다. 특히, 본 제품에 포함된 나노헬릭스만의 buffer는 시료에서 유래된 다양한 PCR inhibitor에 의해 PCR 반응이 저해되는 것을 방지합니다.

 

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.

Storage

• Store below -20 ℃
• 2x Direct qPCR Premix [Green] should be stored in a dark place.

 

Shelf life

• 12 months