HelixAmp™ Direct RT-PCR Kit is designed for the amplification and detection of target RNA directly from whole blood, animal tissues, and plant tissues without any RNA purification processes. The enzyme mix is an optimized blend of thermo reverse transcriptase, antibody-coupled Taq polymerase, and an RNase inhibitor. The 2x buffer mix contains dNTPs, MgCl₂, and a unique buffer system to resist various PCR inhibitors in tissue samples. A uracil-DNA glycosylase and dUTP applied version is also available.
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.
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