Fast First-Strand cDNA Synthesis Mix for RT (with dsDNase) – Rapid Reverse Transcription Kit
6-in-1 Super Mix: Fewer experimental procedures and calculations
One-click copy of information
Instruction manual- Product Description
- Composition
- Precautions
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Product Overview
This product contains a thermostable reverse transcriptase, along with the necessary reaction buffer, RNase inhibitor, dNTPs, Oligo(dT)20VN, random primers, and other key components. It is a 5× ready‑to‑use pre‑mix; when using, simply add dsDNase and its required buffer, the RNA template, and water to achieve a working concentration of 1×, enabling reverse transcription. The resulting cDNA is primarily intended for downstream qPCR assays.
The reverse transcriptase in this product is obtained by genetically engineering the M‑MLV gene to introduce specific functional modifications and expressing it via recombinant E. coli. This enzyme has been engineered to eliminate RNase H activity, exhibits enhanced thermal stability, enables high‑temperature reverse transcription, and mitigates the adverse effects of RNA secondary structure and non‑specific factors on cDNA synthesis.
Storage conditions
Store at 2–8°C; stable for 3 months. Store at –20°C; stable for 2 years. During use, minimize repeated freeze–thaw cycles. Transport with ice packs.
Demo Usage:
Instructions for Use
One-step method (suitable for RNA samples with low genomic DNA content)
Prepare the following reaction solution on ice:
Reagent Usage amount Fast First-Strand cDNA Synthesis Mix 4 μL dsDNase 1 μL RNA template 50 ng~1 μg Nuclease-Free Water Bring up to 20 μL. Note: RNA extracted using the kit is recommended as the template; the RNA extraction kit from Xiantiang Biological is compatible. Briefly centrifuge, vortex to mix thoroughly, and then proceed with the reaction according to the following protocol:
Temperature Time Purpose 37°C 2 min Remove genomic DNA 55℃ 15 min Reverse transcription 85℃ 5 min Terminate reverse transcription Place the prepared cDNA on ice for subsequent experiments. It can be stored at −20°C for short-term use, with a recommended storage period of no more than one week. For long-term storage, aliquot the sample and store it at −80°C.
Two-step method (suitable for RNA samples with high genomic DNA content)
1. Remove genomic DNA
Prepare the following reaction solution on ice:
Reagent Usage amount dsDNase 1 μL 10×dsDNase Buffer 1 μL RNA template 50 ng~1 μg Nuclease-Free Water Top up to 10 μL Note: RNA extracted using the kit is recommended as the template; the RNA extraction kit from Xiantiang Biological is compatible. Briefly centrifuge, vortex to mix thoroughly, and then proceed with the reaction according to the following protocol:
Temperature Time Purpose 37°C 2 min Remove genomic DNA 65℃ 2 min Inactivate dsDNase to terminate the reaction. Note: If the RNA template contains a high proportion of genomic DNA, the 37°C incubation time may be appropriately extended to 5 minutes.
2. First-strand cDNA synthesis
Prepare the following reaction solution on ice:
Reagent Usage amount The reaction product of “Step 1” 10 μL Fast First-Strand cDNA Synthesis Mix 4 μL Nuclease-Free Water Bring up to 20 μL. Note: RNA extracted using the kit is recommended as the template; the RNA extraction kit from Xiantiang Biological is compatible. Briefly centrifuge, vortex to mix thoroughly, and then proceed with the reaction according to the following protocol:
Temperature Time Purpose 50℃ 15 min Reverse transcription 85℃ 5 min Terminate reverse transcription Note: If the target RNA lacks a poly(A) tail, pre-incubate at 25°C for 10 minutes before proceeding with the reaction according to this protocol.
Place the prepared cDNA on ice for subsequent experiments. It can be stored at −20°C for short-term use, with a recommended storage period of no more than one week. For long-term storage, aliquot the sample and store it at −80°C.
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Product Composition
Component Specification (100T) Item number Fast First-Strand cDNA Synthesis Mix 400 μL 500-101A dsDNase 1×100 μL 500-101B 10×dsDNase Buffer 200 μL 500-101C Nuclease-Free Water 2 × 1 mL 500-101D -
Precautions
1. It contains Oligo(dT)20VN and random primers, making it suitable not only for eukaryotic mRNA with a poly(A) tail but also for prokaryotic RNA, eukaryotic rRNA, tRNA, and other templates that lack a poly(A) structure; however, it is not applicable to small RNA templates such as miRNA.
2. Use only after the Mix has been completely thawed and melted, and before use, invert the container several times to mix thoroughly; do not vortex or shake vigorously to avoid excessive bubble formation.
3. Both the purity and integrity of RNA can affect cDNA yield and downstream qPCR results; therefore, use a kit to extract RNA whenever possible.
Keywords
GOONIE
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