Arabidopsis thaliana DNA Extraction in Microcentrifuge Tubes

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Materials in PrecisionPak™

Plant DNA Extraction Kit Contents
Solution LA (Lysing)	60 mL
Solution PA (Precipitation)	15 mL
Solution CA (Clean Up)	30 mL

Add 500 μL of Solution LA into each lysis kit tube.
Add 1-2 cm² or 50 mg of plant material to the buffer-filled tubes.
Note: Cut sample into small pieces – larger sample size may affect complete homogenization with the recommended homogenization setting.
Set the Bullet Blender at speed 12, time 3 minutes and homogenize the samples. If using other homogenizer models, refer to the manufacturer’s instructions for settings.
Remove the tubes and visually inspect the samples to confirm complete homogenization (Figure 1).
Note: If unhomogenized tissue is seen, homogenize for additional 30 seconds.

Remove approximately 500 μL of the homogenate from the lysis tubes and transfer to a new tube containing an equal amount of Solution LA.
Add 100 μL of Solution PA to each tube and mix thoroughly by inversion.
Centrifuge the samples at 10,000 RPM for 5 minutes to pellet the contaminants.

Transfer 750 μL of the supernatant that contains the DNA into new tubes with 750 μL of Solution CA. Mix thoroughly by inversion and incubate at room temperature for 5 minutes.
Centrifuge the solutions at 13,000 RMP for 7 minutes to pellet the DNA.
Remove the supernatant with a 1 mL pipette.
Re-spin the tubes briefly and remove any remaining supernatant with a 200 μL pipette.
Add 30 μL of molecular grade water to the pellet.
Allow the DNA to rehydrate for 10 minutes (about 20% rehydration) or overnight (100% rehydration).
Analyze quality (OD₂₆₀/OD₂₈₀) and yield using a NanoDrop™ or other spectrophotometer and agarose gel (Data of gDNA is shown in Figure 2 and Table 1).
Isolated DNA can be stored at 4°C for up to a week or at -20°C for long term storage.

Table 1. NanoDrop™ Readings.

Tissue	Yield: µg/mg Tissue	OD_260/280
Arabidopsis thaliana 1	0.0100	1.7600
Arabidopsis thaliana 2	0.0860	1.9000