Blood PCR Master Mixes for Serum and Plasma Tests
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Meridian’s Blood PCR Master Mix: Inhibitor Solutions
In addition, PCR inhibitors can be found in the anticoagulants used to stabilize blood samples (e.g. EDTA, citrate or heparin). Traditional methods have relied on removing these inhibitors by DNA or RNA extraction prior to testing, however, these methods are problematic, can cause sample loss and are not 100% effective at removing all the inhibitors.
Meridian’s direct blood master mixes are designed to specifically overcome the inhibitors found in blood samples. As a result, they are capable of highly sensitive detection in multiplex assays, either using extracted DNA or RNA or crudely processed samples. No further optimization is required aside from the addition of primers and probes.
- Ultra-sensitive detection down to less than 10 copies of DNA or RNA using direct amplification protocols.
- Inhibitor-tolerant to PCR inhibitors found in whole blood, serum, plasma (20%) and anticoagulants (e.g. 20% EDTA).
- Can be used in a liquid or dry format, reducing the cost and complexity of creating ambient-temperature stable assays.
Meridian offers ready to use blood, 4x concentrated, glycerol-free master mixes for PCR and isothermal amplification, that have been formulated for direct use with crude lysates that can be used in a wet format or lyophilized and air-dried to create fast, ultra-sensitive ambient-temperature stable assays.
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FAQ’s
Immunoglobulin G is the major inhibitor of qPCR or LAMP in blood, as it can bind to single-stranded genomic DNA, leading to increased Ct values or time to results. The other two main inhibitors in blood are hemoglobin and lactoferrin that affect the polymerase activity and thus lower the amplification efficiency. Hemoglobin and hematin can also cause some fluorescence quenching, particularly for FAM, although this should not affect the speed or Ct value. In addition, inhibitors can be found in the anticoagulants used to stabilize blood samples (e.g. EDTA, citrate, or heparin).
We have used up to 20% whole blood with our blood master mixes, higher concentrations cause the solution to become viscous and difficult to work with.
Yes, the blood master mixes can be used to detect both cfDNA and cfRNA, making it ideal for liquid biopsy.
We do not see a performance difference between the two master mixes.
With Meridian’s direct blood master mixes, you can see down to less than 10 copies.
They are compatible with standard cycling conditions and can also be used with very fast cycling conditions, however some assay development may be required for the volume of blood that is needed for a molecular assay.
It is stable for up to 2 years at ambient temperature if correctly stored in sealed pouches.
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