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Cat. No.: SBDPH-100 (for 100T)

Cat. No.: SBDPH-500 (for 500T)

Description

Simple-Load™ Blood Direct PCR Master Mix (2X) is a conveniently designed PCR pre-mix with a 2X concentration, tailored for blood samples. Following PCR amplification, samples can be directly loaded onto electrophoresis gels without the need for additional loading buffer. This product features the robust BloodTaq™ DNA polymerase, engineered for blood tolerance. BloodTaq™ DNA polymerase is highly efficient and suitable for direct PCR testing of blood samples with anticoagulants such as EDTA, heparin, or sodium citrate, as well as dried blood spots.

Features

• Direct PCR testing of whole blood samples is feasible without the need for DNA extraction. This product demonstrates exceptional resistance to various PCR inhibitors, including hemoglobin, present in human, mouse, and other blood samples. It is suitable for fresh, 4°C stored, or cold-preserved human or mouse blood samples with EDTA, heparin, or sodium citrate anticoagulants, as well as dried blood spots (such as dried blood spots on Whatman 903 or FTA cards), eliminating the requirement for DNA extraction or purification and allowing direct PCR amplification of target DNA.
• Minimal blood samples are required, and the product exhibits high blood tolerance. When used in a 20 μl PCR amplification system, typically around 1 μl of anticoagulated whole blood is sufficient for successful PCR detection. For detecting microbial infections such as bacteria or viruses, to achieve the highest detection sensitivity, up to 4 μl of blood (20% of the volume) can be added to a 20 μl PCR amplification system. For certain anticoagulated blood types (e.g., human heparinized blood), the maximum blood volume that can be added can reach 10 μl (50% of the volume).
• The kit is compatible with three common anticoagulants: EDTA, heparin, and sodium citrate. When using blood samples for PCR testing, all three anticoagulants are compatible. For optimal results, EDTA anticoagulated blood is recommended as it can chelate various metal ions necessary for enzymatic activity, thus better inhibiting nucleic acid degradation.
• The kit provides a pair of positive control primers for easy confirmation of PCR testing efficacy. These primers are universal for mammals and can amplify a 237 bp DNA segment from the upstream non-coding region of the SOX21 gene, suitable for most vertebrates.
• The blood tolerance volume percentage of this product can range from 20% to 50%. The product exhibits blood tolerance percentages of 50%, 20%, and 40% for heparin, sodium citrate, and EDTA-treated human blood, respectively. For PCR testing of mouse blood, the product can tolerate a blood volume percentage of no less than 20%. In conclusion, when using this product for PCR testing, the recommended blood usage rate is 1-20% of the total PCR volume, with a prioritized usage rate of 5-10%.
• The high-fidelity of the BloodTaq™ HF DNA Polymerase (Blood-resistant) contained in this product surpasses that of Pfu DNA Polymerase. As a result, this product can not only be used for blood-related testing purposes but also for PCR amplification and cloning of specific genes.
• The PCR products obtained from this product have blunt ends and cannot be directly used for TA cloning by ligation with T vectors.
• This product can easily amplify target fragments of 5 kb or longer.

Storage

Store at -20°C. Avoid repeated freeze-thaw cycles.

Precaution

• Due to the high sensitivity of PCR amplification, when using the one-step genotyping assay kit for rodents, be cautious to prevent contamination of trace amounts of DNA to be amplified. Consider setting up negative controls without template DNA to confirm the absence of DNA contamination.
• The recommended usage of anticoagulated blood is around 5% of the total PCR system volume and can be directly added to the PCR system without any additional processing.
• After the completion of the PCR reaction, it is advisable to centrifuge at 3000-5000g for 3-5 minutes to pellet blood cell debris, facilitating the collection of the supernatant for gel electrophoresis analysis and similar purposes. Note: Due to properties of the anticoagulated blood itself, a transparent gel-like substance might appear at the bottom of the PCR tube after PCR, which is normal.
• Nuclease-Free Water needs to be provided separately.
• This product is intended for scientific research by professionals only and must not be used for clinical diagnosis or treatment. It is not for use in food or drugs and should not be stored in regular households.
• For your safety and health, please wear appropriate laboratory attire and disposable gloves while handling this product.

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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