Products
Molecular Biology Reagents | Consumables
Nucleic Acid Amplification
Reverse Transcription Series
SweScript All-in-One Blue RT SuperMix for qPCR (One-Step gDNA Remover), (for Reverse transcription series)
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Categories : Biochemicals , 1. Chemical and Reagents , Buffers , Servicebio ,
Brand : Servicebio
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Product Information
Product Name | Cat. No. | Spec. |
SweScript All-in-One Blue RT SuperMix for qPCR (One-Step gDNA Remover) | G3329-50 | 50 rxns |
G3329-100 | 100 rxns |
Description/Introduction
The product includes a 5×ready-to-use premix (5×SweScript All-in-One Blue RT SuperMix for qPCR) and gDNA Remover reagent specially developed for the first chain cDNA synthesis, which can remove the genomic residue of template while reverse transcription is performed. In addition, it is equipped with 5×SweScript All-in-One Blue No-RT Control SuperMix for qPCR, as a control without reverse transcriptase to determine whether the qPCR template comes from cDNA. The premixed solution contains all the reagents needed for reverse transcription reaction (SweScript RT, RNase Inhibitor, Random Primer, Oligo (dT)18 Primer, dNTPs and optimized buffer), as well as blue tracer dyes, which helps to reduce sampling errors and provides color indication and tracer for subsequent qPCR. SweScript RT is
a mutant reverse transcriptase obtained by evolutionary screening in vitro, which has the characteristics of high thermal stability and strong ability of continuous synthesis. gDNA Remover contains a heat-sensitive DNase, which is 30 times more active than DNase, acts only on dsDNA and is easily inactivated at high temperatures.
Storage and Handling Conditions
Ship with wet ice, store at -20°C; valid for 12 months.
Product Contents
Component Number | Component | G3329-50 | G3329-100 |
3329-1 | 5×SweScript All-in-One Blue RT SuperMix for qPCR | 200 µL | 400 µL |
3329-2 | gDNA Remover | 50 µL | 100 µL |
3329-3 | Nuclease-free Water | 1 mL | 1 mL |
3329-4 | 5×SweScript All-in-One Blue No-RT Control SuperMix for qPCR | 20 µL | 40 µL |
Manual | One copy | One copy | |
Assay Protocol/Procedures
1. First strand cDNA synthesis and gDNA removal
(1) Prepare the reaction mixture (recommended 20 μL reaction system)
Component | Volume |
5×SweScript All-in-One Blue RT SuperMix for qPCR | 4 μL |
gDNA Remover | 1 μL |
Total RNA/mRNA | 0.1 ng-5 μg/10 pg-0.5 μg |
Nuclease-free Water | Add to 20 μL |
(2) Mix gently and centrifuge.
(3) Reverse transcription program setup:
Temperature | Time |
25°C | 5 min |
42°C | 15~30 min |
85°C | 5 s |
Note
1. Please wear disposable gloves during operation to avoid RNase contamination.
2. Be sure to put the components G3329-1, G3329-2 and G3329-4 on the ice.
1. Components such as G3329-1, G3329-2 and G3329-4 contain high concentration of glycerol and should be briefly centrifuged before use.
3. For high GC or complex templates, the RNA template, nuclease-free water can be premixed and held at 65°C for 5 min, then quickly cooled on ice. Then add other reaction components.
4. For negative control, it is only necessary to replace the equal volume of G3329-1 with G3329-4 in the reaction system.
5. The reverse transcription products can be preserved at -20°C for a short time. If long-term preservation is needed, it is recommended that the products be stored at-80 after sub-packaging, to avoid repeated freezing and thawing.
6. If the subsequent qPCR primers are designed across exons, the steps of genome removal can be omitted.
For Research Use Only!
