This kit includes a rapid tissue lysis reagent and a PCR amplification master mix, enabling direct lysis of mouse tissue samples (e.g., tail, ear, toe, liver, kidney, etc.) to quickly release genomic DNA. It is also compatible with blood and cell suspension samples. The supernatant from the lysate can be directly added to the PCR reaction system without the need for DNA purification, offering a convenient workflow.

Moreover, the kit requires only minimal sample input: 2–5 mm² of ear or liver tissue, 1–5 mm of tail, or 1–2 toes are sufficient for successful amplification.

The included 2× HieffTM Tissue Direct PCR Mix (With Dye) is a ready-to-use 2× concentrated PCR master mix containing all components required for PCR amplification except template DNA and primers. This greatly simplifies the procedure and reduces the risk of contamination. The mix supports robust amplification of DNA fragments up to 10 kb in length.

This kit is ideal for applications such as transgenic screening and mouse genotyping.

Features

Long Amplicon Capability: Supports direct amplification of DNA fragments up to 10 kb.

Rapid Extension: Fast extension rate of 5–10 seconds per kb, significantly shortening PCR run time.

High Inhibitor Tolerance: Compatible with up to 20% cell lysate in the final reaction mix.

Flexible Lysis Options: Efficient sample lysis in just 3 min at 95°C or 15 min at 60°C.

Multiplex Ready: Enables robust 5–7 plex amplification in a single reaction.

Stable & Reliable: Both the kit reagents and lysates demonstrate excellent stability for consistent performance.

Components

Storage

Unopened kit: Store at –25°C ~ –15°C. Stable for 12 months.

After opening:

Components A and B: Store at 4°C. Stable for 12 months. For frequent long-term use, aliquot to avoid cross-contamination.

Component C: Store at –25°C ~ –15°C. Avoid repeated freeze-thaw cycles. Stable for 12 months.

Application

mouse genotyping; transgenic screening; direct amplification PCR.

Figures

1. Long Amplicon Capability: Up to 10 kb.

Figure 1. Supports Direct Mouse PCR up to 10 kb

Mouse tail, ear, toe, and kidney tissues were lysed, and 1 μL of lysate was used to amplify 1–10 kb fragments. Yeasen 10191ES employed a rapid lysis protocol (95 °C for 3 min) and fast extension (5–10 sec/kb), while other brands followed their respective manuals. Results show that even under rapid lysis and fast cycling, Yeasen-10191ES reliably amplifies fragments up to 10 kb.

2. Robust Amplification Across 30–70% GC Content

Figure 2. Robust Amplification Across 30–70% GC Content

Using 1 μL of mouse lysate, 1–10 kb fragments with varying GC content were amplified. Results indicate that 10191ES supports efficient direct PCR of fragments up to 10 kb across a GC range of 30%–70%.

3. Inhibitor tolerance

Figure 3. Excellent PCR Tolerance to Crude Lysate

Different volumes of lysate prepared at 95 °C for 3 min were added to 20 μL PCR reactions, followed by amplification of 1–10 kb fragments at 5–10 sec/kb extension. Data show strong tolerance of PCR components to crude lysate, with lysate input up to 20% of the reaction volume.

4. Compatible with Multiplex PCR

Figure 4. Multiplex amplification performance

Lysates prepared at 95 °C for 3 min were used for 7-plex amplification in 25 μL PCR reactions with a 5 sec/kb extension rate. Results demonstrate that 10191ES enables multiplex amplification of seven targets ranging from 500 bp to 5 kb.

5. Stable Lysate for Long-Term Storage

Figure 5. Stability testing of lysate.

Mouse tail lysates were stored at 25 °C or 4 °C for four weeks and used to amplify 1–10 kb fragments. Compared with freshly prepared lysates, stored lysates still supported successful amplification, indicating good stability.

6. Strong stability

Figure 6. Excellent Kit Stability

The complete 10191ES kit was stored for four weeks at 25 °C or 4 °C, and for two weeks at 37 °C. Amplification of 1–10 kb fragments confirmed that the kit maintains high stability and reliable performance after storage.

Documents:

Safety Data Sheet

10191_MSDS_HB251121_EN.PDF

Manuals

10191_Manual_Ver.EN20251121.pdf