TaKaRa MiniBEST Universal Genomic DNA Extraction Kit

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	The sample may occasionally be taken out from water bath and vortexed to accelerate tissue lysis.

Protocol tips
The sample may occasionally be taken out from water bath and vortexed to accelerate tissue lysis.

Publication protocol

To investigate the effects of RGV-63R and RGV-91R on RGV replication in cells from different species, the DNA level of RGV in EPCs or GSTCs expressing one protein alone or coexpressing both proteins were measured by qPCR, respectively. A total of 2 × 105 EPCs or GSTCs grown in 24-well plates were transfected with 0.5 μg of empty vector pcDNA3.1 or cotransfected with 0.25 μg of pcDNA3.1-63R-3Flag and 0.25 μg of pcDNA3.1, 0.25 μg of pcDNA3.1-91R-HA and 0.25 μg of pcDNA3.1, or 0.25 μg of pcDNA3.1-63R-3Flag and 0.25 μg of pcDNA3.1-91R-HA. The transfected cells at 24 hpt were harvested and subjected to Western blot analysis. For qPCR, at 24 hpt, the transfected cells were infected with RGV with an MOI of 0.5. The infected cells at 48 h post-infection (hpi) were harvested and washed with PBS three times. The viral genomic DNA was then extracted according to the instructions of TaKaRa MiniBEST Universal Genomic DNA Extraction Kit. Each DNA sample was dissolved in 100 μL of elution buffer, and 1 μL of each DNA sample was used for each qPCR reaction. Specific primers MCP-F/MCP-R for RGV MCP gene was used for qPCR. qPCR was carried out using Fast SYBR Green Master Mix with the StepOneTM Real-Time PCR System (Applied Biosystems, Foster City, CA, USA), as described previously [32].

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Manufacturer protocol

Download the product protocol from Takara Bio Inc for TaKaRa MiniBEST Universal Genomic DNA Extraction Kit below.

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