TaKaRa MiniBEST Universal Genomic DNA Extraction Kit

Protocol tips

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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

Measurement of mtDNA content
Genomic DNA used for the analysis of mtDNA content was isolated from SH-SY5Y cells using TaKaRa MiniBEST Universal Genomic DNA Extraction kit (TaKaRa Bio Inc.). The relative mtDNA copy number was determined by qPCR with primers for the mitochondrial 16S rRNA gene and the nuclear Actin gene. PCR amplification was performed with the qTOWER 2.0 Real Time PCR Thermal Cycler (Analytik Jena) in 20 μL volume containing 50 nM of each primer, 10 μL SYBR Green PCR Master Mix for detection and 20 ng DNA as template. All PCR reactions were performed in triplicates. Primers used were: 5′-GGTGCAGCCGCTATTAAAGG-3′ (16S rRNA, forward); 5′-ATCATTTACGGGGGAAGGCG-3′ (16S rRNA, reverse); 5′-TGACGTGGACATCCGCAAAG-3′ (ACTB, forward); 5′-CTGGAAGGTGGACAGCGAGG-3′ (ACTB, reverse).

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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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