HelixAmp™ Direct RT-PCR Kit 는 RNA 정제 과정 없이 혈액, 동물 및 식물 조직으로부터 직접 target RNA를 증폭/검출할 수 있는 제품입니다. 제품에 포함된 Enzyme Mix에는 antibody-inhibited hot start PCR 효소인 Ab+Taq polymerase 와 함께 Thermo Reverse Transcriptase, 그리고 RNase Inhibitor 등이 최적화된 조건으로 구성되어 있으며, 2x Buffer Mix에는 dNTPs, MgCl₂ 및 나노헬릭스만의 독특한 buffer system이 적용되어 다양한 PCR inhibitor들의 영향을 최소화하였습니다. Carryover contaminant DNA에 의한 오염 방지를 위해 UDG와 dUTP가 적용된 제품을 선택할 수 있습니다.
Figure 1. Comparison of multiple detection for swine influenza virus by direct RT-PCR with conventional RT-PCR from cultured virus.
Lysate of influenza viral particles were applied into direct RT-PCR for multiple detection. Efficiency of detection sensitivity for direct RT-PCR was compared with conventional RT-PCR using total RNAs purified from the same amount of viral particles. NTC: Negative control.
Figure 2. Comparison of amplification efficiency of direct RT-PCR with conventional RT-PCR from whole blood (A) or buccal swab (B).
Lysates of each sample were used for direct RT-PCR as indicated for the volume above. For the reaction comparison, the same amount of each sample was used in total RNA purification and the purified total RNAs were applied in conventional RT-PCR as indicated in the volume above. NTC: Negative control.
Figure 3. Comparison of detection sensitivity of viral target in direct RT-PCR with conventional RT-PCR from animal RNA virus-spiked whole blood.
Lysate of blood spiked with virus particles were used for direct RT-PCR. Total RNAs purified from the same amount of virus-spiked blood were used for conventional RT-PCR. NTC: Negative control.
Figure 4. Comparison of detection sensitivity of direct RT-PCR with conventional RT-PCR from virus-infected plant seeds.
Melon necrotic spot virus(MNSV)-infected melon seeds were used in this experiment. After mixing a grain of crushed seed with a dilution buffer, lysate was used for direct RT-PCR. Total RNAs purified from the same amount of MNSV-infected plant seed were used for conventional RT-PCR. NTC: Negative control.
