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Cat. No.: OGAKR-100 (for 100T)

Cat. No.: OGAKR-500 (for 500T)

Description

One-step Genotyping Assay Kit for Rodents is a particularly convenient kit for rapid genotyping of rodents using a one-step PCR approach directly from blood samples. This kit eliminates the need for genomic DNA extraction, as blood samples can be directly used in PCR. Furthermore, after PCR amplification, samples can be directly loaded onto electrophoresis gels without requiring the addition of loading buffer.

This kit employs 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. This ensures that the kit can be used swiftly and conveniently for genotyping rodents.

Features

• For whole blood samples, direct PCR testing is possible without the need for DNA extraction. This product exhibits exceptional resistance to various PCR inhibitors present in whole blood, such as hemoglobin, for mice, rats, and more. Freshly collected, 4°C stored, or cold-preserved EDTA, heparin, or sodium citrate anticoagulated human or mouse blood samples, as well as dried blood spots (e.g., dried blood spots on Whatman 903 or FTA cards), do not require DNA extraction or purification and can be directly used for PCR amplification of target DNA.
• Minimal blood samples are required, and the product demonstrates 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. While a maximum of 4 μl of blood can be added to a 20 μl PCR amplification system (20% volume), it is generally recommended to use 1 μl of blood sample.
• 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 mice, rats, and humans, capable of amplifying a 300 bp DNA segment of the β-actin gene in mouse, rat, or human genomic DNA.
• The blood tolerance volume percentage of this product can range from 20% to 50%, but a blood usage rate of 5% is recommended. The product exhibits blood tolerance percentages of 50%, 20%, and 30% for heparin, sodium citrate, and EDTA-treated whole blood, respectively. Despite its robust blood tolerance, a recommended blood usage rate of 5% is prioritized for practical use. However, due to factors such as hyperlipidemia, actual blood usage might increase, leading to a potential risk of reduced PCR efficiency.
• The PCR products obtained from this product carry 3'-dA overhangs, which are sticky ends and can be directly used for TA cloning by ligating with T vectors.
• This product can effortlessly amplify target fragments of up to 5 kb in length.

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.

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

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