Rabbit blood DNA – QIAamp DNA blood Midi kit
Extraction Method
Rabbit blood DNA – QIAamp DNA blood Midi kit
FOR RESEARCH USE ONLY
Contents
Materials
Method
Results
Sequencing performance
Change log
Materials
- 1 ml of rabbit blood
- QIAamp DNA blood Midi kit
- TE buffer (1 mM EDTA, pH 8.0)
- 15 ml Falcon tubes
- Centrifuge capable of taking 15 ml Falcon tubes
- 96-100% ethanol
Method
- Lyse the cells using the reagents and volumes according to the Qiagen QIAamp DNA blood Midi kit protocol for volumes of blood between 1–2 ml (page 19, steps 1–3).
- Critical step: We have found that lysis for 1 hour at 60°C is more effective that the recommended 10 minute incubation at 70°C (step 4).
- Purify the lysate according to the protocol (page 20-21, steps 5-11).
- Critical step: To maximize the DNA yield, we recommend adding 100 µl of TE (1 mM EDTA, pH 8.0) to the membrane followed by incubation at 37°C for 10 minutes. Spin the column at 8000 rpm for 2 minutes, and collect the eluted DNA. Repeat the above step twice to give a total elution volume of 300 µl.
- Take 60 µl of eluate (corresponding to 3 µg of DNA) and perform a SPRI size selection.
Results
- Yield: 12-16 μg (before SPRI size selection)
- OD 260/280: 1.96 (after SPRI size selection)
- OD 260/230: 2.33 (after SPRI size selection)
- Fragment size (FEMTO pulse):
Sequencing performance
Libraries were prepared using the Ligation Sequencing Kit and Rapid Sequencing Kit.
- Read length profile:
Change log
| Version | Change |
|---|---|
| v1, 27th July 2019 | Initial protocol publication |
| v2, 14th August 2023 | Updated protocol page references |
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