ARCTURUS® PicoPure® DNA Extraction Kit

Protocol tips

Upstream tips	Protocol tips	Downstream tips
	Use proteinase K extraction solution as quickly as possible following the reconstitution and discard any unused solution

Protocol tips
Use proteinase K extraction solution as quickly as possible following the reconstitution and discard any unused solution

Publication protocol

Timed pregnancy analysis and genotyping
For timed pregnancy analysis, heterozygous females were mated with heterozygous males with or without hormonal stimulation. At day 2.5 or 3.5 dpc, females were sacrificed and the embryos were flushed out of the uterus. Genomic DNA was isolated by using blastocyst lysis buffer (10 mM Tris-HCl, pH 8.5, 50 mM KCl, 2 mM MgCl2, 0.45% NP-40, 0.45% Tween 20, 60 µg/ml proteinase K) and subjected to multiplex PCR analysis with or without genomic DNA amplification by using GenomiPhi DNA Amplification kit (GE Healthcare, Piscataway, NJ) according to the manufacturer's instruction. For later gestation time points, the embryos were fixed in 4% paraformaldehyde, rinsed under tap water, stored in 70% EtOH and paraffin-embedded for sectioning onto metal LCM slides. LCM slides were deparaffinized 5 min in xylene and rehydrated in 100% EtOH for 2 min, 95% EtOH for 2 min, 70% EtOH for 2 min, 50% EtOH for 2 min and dH2O for 2 min. Slides were stained with methyl green for 30 sec and washed in dH2O for 1–5 min. Finally slides were air dried and a portion of embryo was microdissected into PCR caps containing 20 µl of digestion buffer (Arcturus DNA Extraction Kit). Tissue was digested overnight as suggested by Arcturus protocol. A 5 µl of digest was used per PCR reaction by using Advantage 2 PCR kit (Clontech, Palo Alto, CA).

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Manufacturer protocol

Download the product protocol from Thermo Fisher Scientific for ARCTURUS® PicoPure® DNA Extraction Kit below.

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