AcuGenix™ Fast Direct RT-qPCR Master Mix with UNG-V4
AcuGenix™ Fast Direct RT-qPCR Master Mix with UNG-V4, MD2084-4, is designed to perform PCR directly from samples without RNA extraction or sample preparation.
Product overview
AcuGenix™ Fast Direct RT-qPCR Master Mix with UNG-V4 is designed to perform PCR directly from samples without RNA extraction or sample preparation. This reagent consists of rapid DNA polymerase, Neoscript RTase, heat-labile uracil DNA glycosylase (TS-UNG), RNase Inhibitor, MgCl2, dNTPs (with dUTP instead of dTTP), and stabilizers, for one-step reverse transcription and quantitative PCR (qRT-PCR). This reagent has high inhibitor tolerance, and thus it can be directly applied to the detection of samples like throat swab, saliva, anti-coagulated whole blood, plasma, and serum without RNA extraction. This reagent uses special buffer for qPCR with mixed enzymes of anti-inhibitory DNA polymerase and TS-UNG. Therefore, it can obtain a good amplification of target genes in samples containing inhibitors and prevent false positive amplification caused by PCR residual and aerosol pollution.
Available versions
| Cat. no. | Product name | Formulation | Residual DNA level |
|---|---|---|---|
| MD2084-4 | AcuGenix™ Fast Direct RT-qPCR Master Mix with UNG-V4 | Liquid (with glycerol) | Standard |
| FMD5084-4 | AcuGenix™ Fast Direct Lyo-ready RT-qPCR Master Mix with UNG-V4 | Lyo-ready | Standard |
| MD2084-4-NA | AcuGenix™ Fast Direct RT-qPCR Master Mix with UNG (Low DNA)-V4 | Liquid (with glycerol) | Low Residual DNA |
Features and applications
- Reverse transcriptase 42°C~55°C.
- Antibody modified Taq, 95°C, 30s-1min hot start.
- High detection sensitivity.
- Suitable for multiplex multi-channel RT-qPCR.
- Suitable for high-sensitivity amplification of low-concentration RNA templates.
- Suitable for direct sample amplification.
Components
| Component | Specification/Notes |
|---|---|
| 10×Fast Direct RTase/UNG Mix Ⅳ | 10× |
| 2×Fast Direct RT Premix Buffer Ⅳ (dUTP) | 2×, contains dNTPs |
Performance data
1. Freeze-Thaw Test
| 0 freeze-thaw cycles 5 freeze-thaw cycles 10 freeze-thaw cycles 15 freeze-thaw cycles 20 freeze-thaw cycles | |||
| Channel | VIC | ROX | CY5 |
|---|---|---|---|
| Amplification Curves |  |  |  |
Conclusion 1: After 5, 10, 15, and 20 freeze-thaw cycles, the MD2084-4 was consistent with that of reagent not subjected to freeze-thaw cycling, demonstrating stability.
2. Accelerated Stability Test
| MD2084-4 Accelerated at 37°C for 7 d | |||
| Channel | VIC | ROX | CY5 |
|---|---|---|---|
| Amplification Curves |  |  |  |
| MD2084-4 MD2084-4, accelerated at 37°C for 14 d | |||
| Channel | VIC | ROX | CY5 |
| Amplification Curves |  |  |  |
Conclusion 1: After accelerated storage at 37°C for 7 and 14 days, the MD2084-4 was consistent with that of the non-accelerated reagent, demonstrating stability.
3. Rapid PCR Program Test
| Rapid PCR Program: 50°C, 5 min; 95°C, 1 min; 50 cycles (95°C, 1 s; 60°C, 10 s). Respiratory Triple Assay System | |||
| Channel | VIC | ROX | CY5 |
|---|---|---|---|
| Amplification Curves |  |  |  |
| HCV-MLN Plasma Assay System | |||
| Channel | ROX | CY5 | |
| Amplification Curves |  |  | |
Conclusion 1: Under the rapid amplification program, MD2084-4 detected the respiratory pseudovirus extracted template, HCV-MLN Plasma template, and is suitable for rapid amplification.
4. Oral Swab Direct-Amplification Test
| Swab each side of the inner oral wall 10 times, elute in 1 mL H2O, and centrifuge at 2500 r/min for 10 min. | ||
| Channel | Sample Input: 5 μL/T | Sample Input: 10 μL/T |
|---|---|---|
| Amplification Curves |  |  |
| Amplification System | Channel | Concentration | Mean Ct Value |
|---|---|---|---|
| MD2084-4 | VIC | 5μL/T | 24.33 |
| 10μL/T | 23.78 | ||
| ROX | 5μL/T | 31.76 | |
| 10μL/T | 31.60 | ||
| CY5 | 5μL/T | 24.13 | |
| 10μL/T | 23.42 |
Conclusion 1: MD2084-4 can directly amplify human oral swab samples.
Shipping and storage
| Condition | Recommendation |
|---|---|
| Shipping Conditions | -20±5°C |
| Storage Conditions | -20±5°C |
| Shelf Life | 18M |
Downloads
AcuGenix™ Fast Direct RT-qPCR Master Mix with UNG-V4 (MD2084-4)
Instruction Manual (PDF)COA (PDF)MSDS 1 (PDF)MSDS 2 (PDF)AcuGenix™ Fast Direct Lyo-ready RT-qPCR Master Mix with UNG-V4 (FMD5084-4)
Instruction Manual (PDF)COA (PDF)MSDS 1 (PDF)MSDS 2 (PDF)MSDS 3 (PDF)AcuGenix™ Fast Direct RT-qPCR Master Mix with UNG (Low DNA)-V4 (MD2084-4-NA)
Instruction Manual (PDF)COA (PDF)MSDS 1 (PDF)MSDS 2 (PDF)
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