Sarasins minnow brain DNA
Extraction Method
Sarasins minnow brain DNA
FOR RESEARCH USE ONLY
Contents
Materials
Method
Results
Sequencing performance
Materials
- Oryzias sarasinorum brain
- QIAGEN Blood and Cell Culture DNA Midi Kit
- 50 ml Falcon tubes
- Liquid nitrogen
- RNase A
- 70% ethanol
- Isopropanol
- TE buffer (10 mM Tris-HCl, 1 mM EDTA, pH 8.0)
- Agencourt AMPure XP Beads (optional)
- Mortar and pestle
- Incubator or water bath at 50°C
- Centrifuge with capacity for 50 ml Falcon tubes
Method
Snap freeze the sample in liquid nitrogen.
Grind the frozen sample using a pre-cooled (e.g. stored in the freezer for 30 min) mortar and pestle.
Lyse up to 100 mg of the ground, frozen sample according to the standard protocol (steps 3B, 4B, and 5, pages 35-36).
Purify the lysate (steps 1 to 6, pages 49-52).
Critical Step: To maximize the DNA yield, we recommend eluting the DNA overnight in TE buffer (10 mM Tris-HCl, 1 mM EDTA, pH 8.0).
Results
- Yield: 13.7 µg
- OD 260/280: 1.76
- OD 260/230: 0.30
- Fragment size (FEMTO pulse):
Sequencing performance
Libraries were prepared using the Ligation Sequencing Kit.
- Read length profile:
Oxford Nanopore Technologies, the Wheel icon, AmPORE-TB, EPI2ME, GridION, MinION, MinKNOW, PromethION, P2 Solo, and P2 are registered trademarks or the subject of trademark applications of Oxford Nanopore Technologies plc in various countries. Information contained herein may be protected by copyright, patents or patents pending of Oxford Nanopore Technologies plc. All other brands and names contained are the property of their respective owners. Oxford Nanopore Technologies products are RUO. Products labelled/branded as Oxford Nanopore Diagnostics may be RUO or may be regulated as in‐vitro diagnostic devices in some jurisdictions, please check individual product labelling. ONT plc is a member of the producer compliance scheme run by ERP UK Ltd, who manage the submission of documentation in support of WEEE compliance for ONT plc’s manufacture and supply of Electrical and Electronic equipment in the UK. ONT’s WEEE PRN is WEE/MM3828AA.
