Abbexa's Taq Blood PCR Kit consists of a ready-to-use 2X reaction mix specifically designed for fast, highly-specific, direct PCR for whole blood samples. It is suitable for use with samples collected from both human and non-human origins, and with a variety of anticoagulants (EDTA, citrate and heparin). It is developed specifically to overcome PCR inhibitors that are typically present in blood samples. The 250 rxns size is provided as 5 × 625 µl 2X Taq Blood-PCR Mix.

Target	Blood
Tested Applications	PCR
Form	Liquid
Storage	Store at -20 °C. Avoid repeated freeze/thaw cycles.
Validity	Up to 12 months.
Buffer	The exact formulation is proprietary.
Concentration	2X
Availability	Shipped within 3-7 working days.
Note	This product is for research use only. Not for human consumption, cosmetic, therapeutic or diagnostic use.
Directions for use	Reaction Components: Component Volume Whole Blood 1 µl (final concentration 4%) Primers (25 µM each) 0.5 µl Taq Blood-PCR Mix (2X) 12.5 µl Water Variable, up to 25 µl Total Volume 50 µl Thermal Cycling Conditions: Step Number of Cycles Temperature Time per Cycle Initial Denaturation 1 cycle 95 °C 3 min Denaturation 30-40 cycles 95 °C 15 seconds Annealing ≥ 55 °C (primer dependent) 15 seconds Extension 72 °C 45 seconds Notes: Forward and reverse primers are generally used at a final concentration of 0.2-0.6 µM each. It is recommended to start with 0.5 µM as the final concentration. A primer concentration that is too high can reduce the specificity of priming, resulting in non-specific products. Primers should have a melting temperature (Tm) of approximately 60 °C. This kit is suitable for use with whole blood samples collected with various anticoagulants (e.g. EDTA, citrate, heparin). A final whole blood concentration of 4% is recommended, though the kit can be used at concentrations up to 20%. Pipetting following PCR may be difficult if the concentration is above 20%. The concentration may need to be optimised for non-human blood samples. It is recommended to reduce the final whole blood concentration for samples containing nucleated erythrocytes (such as avian blood). The initial denaturation step at 95 °C for 3 minutes is required to activate the enzyme and fully melt the template. It is recommended to run the subsequent denaturation steps at 95 °C for 15 seconds each cycle, which is suitable for GC-rich templates. The optimal annealing temperature is primer dependent and is usually 2-5 °C below the lower Tm of the pair. It is recommended to start with an annealing temperature of 55 °C and, if necessary, run a temperature gradient to determine the optimal annealing temperature. 15 seconds per cycle is usually sufficient, though increasing this step up to 45 seconds may improve problematic reactions. The optimal extension time is dependent on the length of the amplicon and the complexity of the template. An extension time of 15 seconds is sufficient for amplicons under 1 kb. Longer extension times are recommended for fragments larger than 1 kb. The extension time may be increased up to 45 seconds/kb to find the fastest optimal condition. The optimal conditions will vary from reaction to reaction and are dependent on the system used. Each parameter needs to be adjusted by the end user and some optimization may be required.

Research Articles on Blood

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