The Hieff™ Fast Cell Direct SYBR Green RT-qPCR Kit enables direct gene expression analysis from a wide range of animal cells—including adherent and suspension cell lines, primary cells, stem cells, and iPSCs—without prior RNA extraction. This streamlined workflow reduces hands-on time, minimizes handling errors, and delivers results in as little as 1.5 hours, from cell lysis to qPCR readout. The kit includes all necessary reagents for cell lysis, DNase treatment, reverse transcription, and SYBR Green-based real-time PCR, allowing rapid and reliable gene expression profiling.

Features

• Works on all qPCR instruments – ROX dye pre-mixed; no adjustment needed.
• Fast workflow – From cells to results in just 1.5 hours, no RNA extraction required.
• Sensitive & reproducible – Reliable detection for genes with any GC content.
• Tolerant to crude samples – Works with up to 45% cell lysate.
• Highly stable – Store at 4°C for 7 days or freeze/thaw up to 50 times without losing performance.

Components

[Note]: *If needed, additional Lysis set (Cat#16811ES), RT set (Cat#16812ES), and qPCR set (Cat#16813ES) can be purchased separately.

Storage

Store unopened components 11172-A and 11172-B at –25°C to –15°C. After thawing, store at 4°C and avoid contamination. All other components should be stored long-term at –25°C to –15°C. The kit is stable for 6 months when stored as directed.

Application

• Validation of siRNA-mediated gene knockdown;
• Analysis of stimulus-induced gene regulation;
• Drug discovery applications (e.g., target screening and validation)

Figures

1.Universal Compatibility

Figure 1. Instrument platform compatibility testing. Amplification of 293T cell-derived targets using kit #11172ES on multiple qPCR platforms: Bio-Rad (no ROX), ABI QuantStudio 5 (low ROX), and ABI StepOnePlus (high ROX). Results demonstrate consistent performance across instruments with different ROX requirements, confirming universal compatibility of the kit.

2. Excellent gDNA digestion capability.

 Figure 2. gDNA digestion capacity assessment. Genomic DNA (100–500 ng) was spiked into the reaction and treated with either Yeasen’s 11172ES kit or a leading brand kit without DNase I, followed by reverse transcription and qPCR. Results show that 11172ES achieves superior gDNA removal across all tested gDNA input levels compared to the imported product.

3. High sensitivity & repeatability

Figure 3. Sensitivity and repeatability assessment. (A) Suspension and (B) adherent 293 cells were analyzed using Yeasen 11172ES or Supplier T* for genes with 30%, 50%, and 70% GC content. 11172ES consistently delivered lower Ct values, better reproducibility, and higher sensitivity across all GC levels.

4. High Inhibition Tolerance

 Figure 4. High Inhibition Tolerance

a. qPCR Tolerance:Added varying amounts of cell lysate (4 μL [20%], 5 μL [25%], 6 μL [30%], and 9 μL [45%]) to the 11172ES reverse transcription reaction. Results show that 11172ES can tolerate up to 45% cell lysate without loss of performance.

b. qPCR Tolerance: Added different volumes of cDNA (2 μL [10%], 4 μL [20%], and 6 μL [30%]) to the 11172ES qPCR reaction. Results confirm that 11172ES maintains excellent tolerance, handling up to 30% cDNA derived from cell lysates with no significant inhibition.

5. Strong Stability

Figure 5. 293 cells were tested with 11172ES stored at –20°C, 4°C, 25°C, or 37°C for 1–7 days. All conditions showed ΔCt < 0.5, confirming excellent reagent stability under varied storage temperatures.

Documents:

Safety Data Sheet

11172ES-MSDS-HB260109

Manuals

11172-Manual-Ver.EN20260109