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.

Documentation

User Manual

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.

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