Description
Product description
This product is the brand's highly recommended high-performance cDNA synthesis reagent, featuring faster reverse transcription speed, higher yield, and strong resistance to residual inhibitors. It is suitable for reverse transcription reactions of RNA from animal, plant, and microbial sources. Additionally, the included gDNA Digester Mix can digest up to 1000 ng of DNA, effectively preventing contamination from residual DNA in the RNA template and ensuring more reliable downstream results.
The reverse transcription product of this product can be used for downstream qPCR applications. It is recommended to use in combination with Universal Multiplex qPCR Probe Premix (Cat#N132041), Universal qPCR Dye Premix (Cat#N132031), or Traceable qPCR Dye Premix (Cat#N132034, Cat#N132035, Cat#N132036) for high-performance gene expression analysis.
Specifications
|
Catalog Number
|
N132062E
|
N132062S
|
|
Specifications
|
10 T
|
100 T
|
Components
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Catalog Number
|
Component Name
|
N132062E
|
N132062S
|
|
N132062-A
|
cDNA Fast Synthesis SuperMix
|
60 μL
|
600 μL
|
|
N132062-B
|
gDNA Digester Mix
|
20 μL
|
200 μL
|
|
N132062-C
|
RNase-free H2O
|
200 μL
|
2×1 mL
|
Storage
Store at -25 to -15℃. Valid for 1 year.
Notes
1. All steps for adding samples and preparing solutions should be carried out on ice whenever possible.
2. Before use, each component should be thoroughly mixed by inverting up and down, followed by a brief low-speed centrifugation to collect the contents at the bottom of the tube.
3. For your safety and health, please wear a lab coat and disposable gloves when operating.
4. For Research Use Only.
Instructions
1. Reverse Transcription with gDNA Removal Step
1) DNA Digestion
Prepare the following mixture in an RNase-free centrifuge tube and gently mix by pipetting. Incubate at 37°C for 2 minutes.
|
Component
|
Usage Amount
|
|
gDNA Digester Mix
|
2 μL
|
|
Total RNA or mRNA
|
Total RNA：10 pg-2 μg
mRNA：10 pg-500 ng
|
|
RNase-free H2O
|
Up to 14 μL
|
【Note】It is recommended that the input amount of Total RNA does not exceed 2 µg. If the expression level of the target gene is low, the input amount can be increased up to a maximum of 5 µg.
2) Reverse Transcription Reaction Setup (Example for a 20 µL Reaction Volume)
|
Component
|
Usage Amount
|
|
The reaction mixture from the previous step
|
14 μL
|
|
cDNA Fast Synthesis SuperMix
|
6 μL
|
3) Reverse Transcription Procedure Settings
|
Temperature
|
Time
|
|
50℃
|
5 min
|
|
85℃
|
5 sec
|
【Note】The reverse transcription product can be directly used for downstream qPCR detection. If the downstream experiment will not be performed immediately, the product can be stored at 4°C. For long-term storage, it is recommended to aliquot and store at -20°C to avoid repeated freeze-thaw cycles.
2. Reverse Transcription without gDNA Removal Step
1) Reverse Transcription Reaction Setup (Example for a 20 µL Reaction Volume)
|
Component
|
Usage Amount
|
|
cDNA Fast Synthesis SuperMix
|
6 μL
|
|
Total RNA or mRNA
|
Total RNA：10 pg-2 μg
mRNA：10 pg-500 ng
|
|
RNase-free H2O
|
Up to 20 μL
|
2) Reverse Transcription Protocol Setup
|
Temperature
|
Time
|
|
50℃
|
5 min
|
|
85℃
|
5 sec
|
【Note】The reverse transcription product can be directly used for downstream qPCR detection. If the downstream experiment will not be performed immediately, the product can be stored at 4°C. For long-term storage, it is recommended to aliquot and store at -20°C to avoid repeated freeze-thaw cycles.
