Blood Direct PCR Kit (with Dye)
Based on 1 Customer Validation
MCE Blood Direct PCR Kit (with Dye) is specifically designed for direct amplification from blood samples. It enables PCR amplification directly from whole blood without the need for conventional DNA extraction and purification steps, thereby significantly simplifying the workflow and improving detection efficiency.
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Storage :
-20°C, 1 year. Shipping with dry ice.
To place orders, for customer services and technical support, please contact: MedChemExpress USA
Tel: 609-228-6898 E-mail: [email protected] [email protected]
Description & Advantages
MCE Blood Direct PCR Kit (with Dye) is specifically designed for direct amplification from blood samples. It enables PCR amplification directly from whole blood without the need for conventional DNA extraction and purification steps, thereby significantly simplifying the workflow and improving detection efficiency.
This kit is compatible with various types of blood samples, including fresh whole blood treated with common anticoagulants such as EDTA, heparin, and citrate, as well as refrigerated or frozen blood samples and commercially available dried blood spot (DBS) cards.
Features of MCE Blood Direct PCR Kit (with Dye):
1. Fast and Convenient: Direct PCR amplification from whole blood without DNA extraction.
2. High specificity and efficiency: Utilizes an optimized Taq polymerase system with strong template affinity, high amplification specificity and fidelity, and robust tolerance to common PCR inhibitors.
3. High amplification performance: Efficient amplification of target fragments up to ≤ 8 kb; strong GC compatibility and rapid extension rate (up to 10 kb/s), significantly reducing experimental time.
4. Broad sample compatibility: Suitable for whole blood samples from different species and treated with various anticoagulants, as well as refrigerated/frozen blood and dried blood spot (DBS) samples.
Protocol
General Protocol
1. PCR reaction mixture
| Component | 50 μL reaction |
|---|---|
| Advanced PCR Buffer | 25 μL |
| Advanced High-Fidelity DNA Polymerase Mix | 1 μL |
| Forward Primer (10 μM) | 2 μL |
| Reverse Primer (10 μM) | 2 μL |
| Blood sample | X μL |
| ddH2O | To 50 μL |
Note: a. Mix all components thoroughly before use;
b. A final concentration of 0.4 μM for each primer is recommended;
c. The optimal whole-blood template concentration is 0.5%-20% of the total reaction volume. A starting condition of 10% is recommended, i.e., 5 μL whole blood in a 50 μL reaction. Avoid aspirating blood clots;
d. For dried blood spots on filter cards, approximately 1 mm2 of blood-stained paper may be placed directly into the PCR reaction without pretreatment.
2. PCR cycling program
| Cycle step | Temperature | Time | Cycles |
|---|---|---|---|
| Initial denaturation | 95°C | 5 min | 1 |
| Denaturation | 95°C | 15 sec | 30-35 |
| Annealing | 60°C | 15 sec | 30-35 |
| Extension | 72°C | 3-10 sec/kb | 30-35 |
| Final extension | 72°C | 5 min | 1 |
Note: a. Use the theoretical primer Tm or 1-2°C below the primer Tm as a starting annealing temperature; use a temperature gradient if specificity is poor;
b. An extension time of 10 sec/kb amplifies most targets below 8 kb, while 3-5 sec/kb is suitable for most targets below 2 kb. If amplification efficiency is low, extend to 20-30 sec/kb and do not exceed 60 sec/kb.
3. PCR product processing
After PCR, centrifuge the reaction product at approximately 4,000 rpm (about 1,000 × g) for 1-3 min to pellet blood-cell debris, and collect the supernatant for downstream analysis. Add MCE DNA loading buffer when agarose gel electrophoresis is required.
Storage
-20°C, 1 year.
Shipping with dry ice.
Attention
1. This product is for R&D use only, not for drug, household, or other uses.
2. For your safety and health, please wear a lab coat and disposable gloves to operate.
Components
| Components | HY-K0537-50 T |
|---|---|
| Advanced PCR Buffer | 1.25 mL |
| Advanced High-Fidelity DNA Polymerase Mix | 50 μL |
| Positive Control Primer Mix (10 μM) | 100 μL |
| DNA loading buffer (10×) | 500 μL |
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