Description

The Extraction-Free 1-Step RT-PCR Master Mix, SYBR Green Dye (#142) enables real-time qualitative and quantitative RT-PCR directly from cultured cells without the need for RNA purification. This 2X premixed solution contains all necessary components for one-step RT-PCR except gene-specific primers and RNA templates.

It streamlines gene expression analysis by allowing direct amplification from crude lysates, eliminating time-consuming and laborious RNA purification steps.

Key Features

One-step RT-qPCR workflow: reverse transcription and PCR amplification combined in a single tube.
Direct-from-sample: enables expression analysis directly from cultured cells without RNA purification.
Fast preparation: 5-minute lysis including DNase treatment, versus 30–60 minutes for traditional RNA purification.
Robust performance: supports 10 to 100,000 cells per sample, results comparable to purified RNA.
Economic and efficient:
2 × 1 mL packaging—200 reactions at 20 μL or 400 reactions at 10 μL final volume.
SYBR Green detection method.
Compatible with most real-time PCR instruments.
Try Before You Buy — Request a trial sample.

Kit Components

2X Master Mix (2 × 1 mL / 5 × 1 mL / 10 mL for 200, 500, or 1000 × 20 μL reactions).
Lysis Buffer (2 × 10 mL / 1 × 50 mL / 1 × 100 mL).
User Manual.

Storage

Store at -20°C (stable up to 24 months with ≤10 freeze-thaw cycles). Transport at ≤4°C for up to 3 days.

Reaction Setup

20 μL Reaction: 10 μL Master Mix, primers, ≤4 μL RNA sample, water to 20 μL.
10 μL Reaction: 5 μL Master Mix, primers, ≤2 μL RNA sample, water to 10 μL.
Recommended amplicon size: ≤150 bp.

Compatible Instruments

Compatible with all major real-time PCR instruments, including but not limited to Applied Biosystems®, Bio-Rad®, Roche®, Qiagen®, Eppendorf®, and more.

Workflow

1. Harvest 10,000–100,000 cells.

2. Add lysis buffer and incubate for 5 minutes.

3. Heat at 95°C for 10 minutes.

4. Add lysate directly to RT-PCR setup.

5. Run RT-qPCR and analyze results.

Product Comparison

142: SYBR Green, No multiplexing, No probe system, Standard single-gene expression.

146: TaqMan, Yes multiplexing, Yes probe system, Multiplex TaqMan assays.

149: TaqMan, Yes (Super Multiplex), Yes probe system, Highly complex multi-target detection.

Documents

User Manual

Extraction-free RT-PCR from cultured cells

FAQ

Why Choose Biofargo #142?

Our Extraction-Free 1-Step RT-PCR Master Mix, SYBR Green Dye (#142), is designed as a cost-effective alternative to Thermo Fisher's Cells-to-CT™ 1-Step Power SYBR™ Green Kit.

With comparable performance in gene expression analysis directly from cell lysates, our kit offers reaction costs as low as $4 per reaction—ideal for both research laboratories and high-throughput applications.

What nucleotides are included in the 2× Probe qPCR Master Mix? Does it contain UTP?

The 2× Probe qPCR Master Mix contains all standard deoxynucleotides—dATP, dTTP, dGTP, and dCTP—each at a standard concentration of 200 µM.
UTP is not included by default, but custom formulations with UTP can be provided on request, depending on the required quantitative range.
Additionally, the mix uses a next-generation hot-start enzyme, which allows for higher reaction temperatures and results in enhanced dNTP stability and overall amplification performance.

Documents

User Manual

Extraction-free RT-PCR from cultured cells

Extraction-Free 1-Step 2X RT-PCR Master Mix, SYBR Green Dye (#142)-Biofargo