Tissue Extract-PCR Buffers
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Meridian's Tissue Extract-PCR Buffers
- Produces high yield, PCR-ready DNA in as little as 15 minutes
- Reduces the risk of sample loss and contamination and minimizes manual effort
- Use with high quality Taq HS DNA Polymerase (MDX008) for improving yield of even the most challenging targets
- No need for hazardous chemicals or multiple washing steps
Sex determination in neonate mice
Mouse (Mus musculus) tail tissue placed into equal volumes of Buffer A and Buffer B and incubated at 75°C for 5 minutes, followed by deactivation at 95°C for 10 minutes. Taq DNA Polymerase (MDX008) and X- and Y- chromosome-specific gene were added, amplified and the PCR products loaded onto an agarose gel. A single band is seen in five of the nine reactions (Lanes 1, 3, 5, 7 and 9) correspond to the XX (female chromosomes) and a doublet in the other four (Lanes 2, 4, 6, and 8) correspond to the XY (male chromosomes). The PCR product sizes were checked and corresponded exactly to a visual determination of the sex of the individual mice.
Tissue Extract-PCR Buffers, MDX004
Convenient, fast and efficient method for the extraction of DNA from a variety of mammalian tissues in a single tube, without the need for multiple washing steps for use directly in a PCR reaction.
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Description
Tissue Extract-PCR Buffer offers a convenient, fast and efficient method for the extraction of DNA from a variety of mammalian tissues, particularly from rodent tail or ear samples. The DNA extractions are performed in a single tube, without the need for multiple washing steps, greatly reducing the risk of sample loss and contamination, and can be used directly in a PCR reaction.
Specifications
| Description | A convenient, fast and efficient method for the extraction of DNA from a variety of mammalian tissues in a single tube, without the need for multiple washing steps for use directly in a PCR reaction. |
| Concentration | 5x |
| Appearance | Clear, colorless solution |
| Application | PCR, RT-PCR, qPCR, RT-qPCR, LAMP |
| Sample type | cDNA, DNA |
| Presentation | 2 vials |
| Storage | -20 °C |
| Mix stability | See outer label |
| Sensitivity | PCR with a dilution series of extracted DNA, to determine specific product at limiting template concentration |
| DNA Contamination | None detected in PCR amplification with traces overlay with the negative control on E. coli and mouse genomic DNA specific targets. |
| DNase Contamination | No detectable degradation |
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FAQs: Tissue Extract-PCR Buffers
Is it critical to deactivate at 95°C for ten minutes?
Yes, if it is not properly deactivated through the heat denaturation step, it could quickly degrade the polymerase during the PCR step.
Can the tissue extract be used for qPCR?
Yes, the lysate can be used directly in a qPCR reaction, we recommend using the inhibitor-tolerant or Specimen-specific™ mixes.
Can the tissue extract be stored?
Yes, the tissue extract can be stored at -20°C in the short term but for long term storage, we recommend purifying the DNA and re-suspending it in a standard buffer.
What sample types can be used with the Tissue Extract-PCR Buffers?
The Tissue Extract-PCR Buffers has been tested with cell cultures, tissues (tail, ear, kidney, liver, blood and FFPE), buccal swabs, stool samples, gram positive bacteria and hair follicles.
Is centrifugation a requirement prior to PCR?
Depending on the sample type, it is possible to omit the centrifugation step after the extraction reaction, however pelleting the debris will result in a more reliable PCR.
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