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Lyo-Ready Direct DNA and RNA/DNA qPCR Blood Master Mixes

Lyo-Ready Direct DNA qPCR and RNA/DNA qPCR Blood mixes are glycerol-free mixes that contain optimized excipients compatible with freezing and lyophilization and they designed for the direct molecular quantitation of DNA and RNA from whole blood, serum, or plasma.

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Meridian's Lyo-Ready Direct DNA and RNA/DNA qPCR Blood Master Mixes

  • Create ultra-sensitive multiplex direct blood qPCR tests using whole blood, serum or plasma.
  • Simplify the development of quantitative PCR blood tests, by removing the nucleic acid extraction step and shortening the sample-to-result workflows.
  • Validated for detection of blood pathogens and liquid biopsy applications.
  • Can be used in liquid format or it can be lyophilized, for developing an ambient-temperate stable assay.

Significantly greater tolerance to inhibitors found in whole blood and anticoagulants

Lyo-Ready Direct DNA qPCR Blood, MDX122

Available in 5 mL (1,000 Rxns) or 50 mL (10,000 Rxns) aliquots

 

Lyo-Ready Direct RNA/DNA qPCR Blood, MDX123

Available in 5 mL (1,000 Rxns) or 50 mL (10,000 Rxns) aliquots

Blood is one of the most common specimens used for laboratory diagnostic testing and it is useful for evaluating the function of vital organs (kidneys, liver, thyroid, and heart) and for diagnosing diseases such as bacterial and viral infections, cancer, cardiovascular disease, and metabolic disorders such as diabetes. However, whole blood specimens, serum, and plasma contain a number of inherent PCR inhibitors including immunoglobulin G, hemoglobin, lactoferrin, and leukocyte DNA. 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.

Lyo-Ready DNA qPCR Blood is a 4x qPCR master mix for the development of DNA blood tests, enabling highly sensitive detection of target DNA from whole blood, plasma, and serum samples. Lyo-Ready RNA/DNA qPCR Saliva is 4x qPCR master mix and can be used for the development of both RNA and DNA blood tests from whole blood, plasma, and serum samples. These mixes have been formulated specifically to overcome the inhibitors specifically found in blood samples – no further optimization is required aside from the addition of primers and probes. Furthermore, these mixes can be used in a wet format or lyophilized to create ambient-temperature stable whole blood tests, plasma tests or serum tests.

FAQs: qPCR Blood Test

Blood contains abundant information about the health status of the organism, it can be used to evaluate organ function, detect proteins, nucleic acids, metabolites, infectious diseases and can be used for diagnosis of cancer (liquid biopsy), heart disease, diabetes. Blood tests are a simple and easily accessible (low-invasive) method to assess and monitor health.

Immunoglobulin G is the major inhibitor of qPCR in blood, as it can bind to single-stranded genomic DNA, leading to increased Ct values in a quantitative PCR blood test. The other two main inhibitors in blood are hemoglobin and lactoferrin that affected the DNA polymerase activity and thus lowered the amplification efficiency. Hemoglobin and hematin can also cause some fluorescence quenching, particularly for FAM, although this should not affect the 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 mix, higher concentrations cause to solution to become viscous and difficult to work with.

We use our own proprietary Taq polymerase and a highly optimized buffer system to overcome inhibition.

It is stable for up to 2 years at ambient temperature if correctly stored in sealed pouches.

Yes, the Lyo-Ready qPCR Blood mixes can be used to detect both cfDNA and cfRNA.

Yes, it is possible that you may need to pre-treat the blood in order to release the DNA from the gram-positive bacteria. In some cases a heat shock is sufficient, this will depend on the bacteria being examined.

We do not see a performance difference between the two mixes, which is why we call the Lyo-Ready Direct RT-qPCR Blood mix – Lyo-Ready Direct RNA/DNA qPCR Blood mix, as it can be used for both.

It can be stored frozen or following the instructions from the storage buffer manufacturer.

They are compatible with standard cycling conditions and can also be used with very fast cycling conditions, however some optimization may be required for the amount of blood that is needed for the qPCR blood test.

You should elute in water, or TE buffer.

Yes, the Lyo-Ready Direct DNA qPCR Blood and Lyo-Ready Direct RNA/DNA qPCR Blood can be used as a liquid mix or lyophilized and stored for up to 24 months without needing to change the reaction conditions and lyophilization/ambient temperature storage will not affect the sensitivity of the test.

We advise not using Triton; however, heat treatment is ok.

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