Biomiga One-Step RT-PCR Kit - RT0201-01

Description :

Introduction

One-Step RT-PCR is designed for the reverse transcription (RT) and polymerase chain reaction (PCR) amplification of a specific target RNA from either total RNA or mRNA. This system uses a mixture of MMLV Reverse Transcriptase and Hot start Taq DNA polymerase in an optimized reaction buffer, and can detect RNA targets up to 4.0 kb. The amount of input total RNA can range from 0.01 pg to 2 μg. Sufficient reagents are provided for 25 or 100 amplification reactions of 50 μl each.

Kit Contents

Storage

Store at -20°C，One-Step RT-PCR is stable for 2 year when stored at -20°C.

Features

• Convenient one-tube setup
• Optimized RT-PCR buffer and robustness
• High sensitivity, specificity, and reproducibility

Recommended RT-PCR reaction assembly

The following protocol is suggested as a starting point.

 * If needed, the magnesium concentration can further be optimized

1. Assemble the reaction on ice.

2. Program the thermal cycler so that cDNA synthesis is followed immediately by PCR amplification.

cDNA synthesis: 1 cycle: 40-50°C for 10 min

Denaturation: 1 cycle: 94°C for 2 min

PCR amplification: 40 cycles:

94°C for 15 s

55-65°C for 30 s

68°C for 1 min/kb

3. Analyze RT-PCR amplified products by gel electrophoresis.