Rabbit blood DNA – QIAGEN Genomic-tip 500/G
Extraction Method
Rabbit blood DNA – QIAGEN Genomic-tip 500/G
FOR RESEARCH USE ONLY
Contents
Materials
Method
Results
Sequencing performance
Change log
Materials
- 10 ml of blood, collected in K2-EDTA
- QIAGEN Blood and Cell Culture DNA Maxi kit
- 70% ethanol in nuclease-free water
- Isopropanol
- TE buffer (1 mM EDTA, pH 8.0)
- 1.5 ml Eppendorf DNA LoBind tubes
- 50 ml Falcon tubes
- Centrifuge and rotor for 50 ml Falcon tubes
- Incubator or water bath set at 50°C
Method
- Perform cell separation and lysis according to the QIAGEN protocol (Preparation of blood samples section) (page 21).
- Critical step: Lyse the nuclei for 1 hour at 50°C to ensure the lysis of all the cells, using the reagent quantities described in the protocol (page 24).
- Purify the lysate according to the QIAGEN protocol (Isolation of Genomic DNA from Blood, Cultured Cells, Tissue, Yeast, or Bacteria using Genomic-tips section) (page 49).
- Critical step: To maximize the DNA yield we recommend that the elution is performed for 2 hours at 50°C, using 200 µl TE (1 mM EDTA, pH 8.0), occasionally mixing the tube contents by gentle inversion.
- Take 3 µg of eluate and perform a SPRI size selection.
Results
- Yield: 90-120 μg
- OD 260/280: 1.93
- OD 260/230: 2.33
Sequencing performance
Libraries were prepared using the Ligation Sequencing Kit.
- Read length profile:
Change log
| Version | Change |
|---|---|
| v1, 27th July 2019 | Initial protocol publication |
| v2, 14th August 2023 | Updated protocol links and page numbers |
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