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Cat. No.: SMPP-100 (for 100T)

Cat. No.: SMPP-500 (for 500T)

Description

Simple-Load™ Multiplex PCR Master Mix (2X) is a highly convenient 2X concentration PCR premix specifically designed for multiplex PCR amplification. After PCR, samples can be directly loaded for electrophoresis without adding a loading buffer. This product contains Multi™ DNA polymerase, which allows for efficient, highly specific, and highly sensitive PCR amplification of multiple (more than 20) target DNA fragments simultaneously and very evenly. Additionally, this product can be used not only for regular multiplex PCR but also for direct PCR of whole blood, serum, or plant samples. This makes it convenient for multiplex PCR detection of bacterial and viral infections, gene mutations in blood samples, or gene mutations and microbial infections in plant samples.

Multiplex PCR is a technique that allows the simultaneous amplification of at least two or more DNA fragments in a single PCR reaction. This technique is widely used in scientific research, disease diagnosis, forensic or diagnostic genotyping, and other fields. It can also be used for quantitative or semi-quantitative expression analysis of genes using cDNA as a template. It is particularly suitable for multi-gene detection in various small samples of animals, plants, fungi, bacteria, or viruses, with outstanding advantages of high specificity and high sensitivity.

Features

• This product exhibits superior multiplex amplification performance. It has been tested to easily achieve efficient, highly specific, and highly sensitive amplification of 15 target DNA fragments simultaneously and very evenly. Even with template amounts as low as 1 pg, it can be well amplified with only 30 PCR cycles. Ordinary PCR kits may show different amplification efficiencies for different primers and templates due to preferences during PCR amplification, leading to some fragments being easily amplified while others are more difficult. This kit uses Multi™ DNA polymerase, which has been carefully screened and mutation-optimized for multiplex PCR, along with repeatedly optimized compatible buffers, ensuring efficient, highly specific, and highly sensitive balanced amplification of multiple target DNA fragments.
• This product is compatible with blood and plant samples. Whole blood or serum samples can be directly used as templates for multiplex PCR detection with this kit without the need for DNA extraction and purification. This product is suitable for EDTA, heparin, or sodium citrate anticoagulated blood samples or dried blood spot samples. Plant samples can also be directly used for multiplex PCR detection with this kit, without the need for DNA extraction and purification.
• This product provides a positive control to facilitate verification of its effectiveness. The kit includes a Control template and primer mix, which is a pre-mixed template and primer that can be used as a positive control to verify the multiplex PCR amplification effect of this product. The Control template and primer mix contains 15 pairs of primers, which can amplify 15 bands ranging from 113 bp to 1463 bp.
• This product is very convenient to use. Simply add primers, templates, and water to perform multiplex PCR, and after PCR, samples can be directly loaded for electrophoresis without adding a loading buffer.
• The amplification products of this product can be used for TA cloning. The PCR products obtained using this product have 3’-dA overhangs, which can be directly used for TA cloning with T vectors. The fidelity of the Multi™ DNA polymerase used in this product is similar to that of Taq DNA polymerase, so this product is mainly recommended for qualitative and semi-quantitative detection.

Storage

Store at -20°C. 

Precautions

• Primer design is crucial for the success of multiplex PCR. Primers should meet conventional design rules to avoid non-specific amplification and failure to amplify. Designed primers should be individually validated by PCR before selecting the best pairs for multiplex PCR. Ideally, the Tm value should be calculated using Tm = 2n(A) + 2n(T) + 4n(C) + 4n(G) (e.g., a primer with 3 A’s, 7 T’s, 4 G’s, and 6 C’s would have Tm = 2×3 + 2×7 + 4×4 + 4×6 = 60). The Tm value should not be lower than 60°C, and typically, a Tm of at least 65°C yields better results for multiplex PCR.
• High-quality primers (desalted, PAGE, or HPLC purified) are recommended. Before use, mix all primer pairs to adjust the stock concentration of each pair to 10 μM, ensuring a final concentration of 0.2 μM in the PCR reaction system.
• The recommended extension rate is 2 min/kb. For difficult-to-amplify targets, the extension time can be adjusted to 3-4 min/kb.
• If there are many PCR product bands, adjust the loading volume for electrophoresis to about 2 μl after the PCR reaction. Excessive loading volume can lead to uneven bands.
• Due to the high sensitivity of multiplex PCR, avoid contamination with trace amounts of DNA to be amplified. It is advisable to set up a blank control without a template to confirm whether there is contamination of the DNA to be amplified.
• Nuclease-Free Water is required.
• For templates with high GC content, it is recommended to use Multiplex PCR Enhancer (2X) in conjunction.
• This product is for scientific research use by professionals only and is not intended for clinical diagnosis or treatment, food or drug use, or storage in ordinary residences.
• For your safety and health, please wear a lab coat and disposable gloves while handling.

Related:

• Simple-Load™ Multiplex PCR Master Mix (2X)
• Simple-Load™ Plant Direct PCR Master Mix (2X)
• Simple-Load™ Blood Direct PCR Master Mix (2X)
• Simple-Load™ Blood Direct PCR Master Mix (HF, 2X)

Only for research and not intended for treatment of humans or animals

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