Genomic DNA Isolation Kit (Plant)

The Genomic DNA Isolation Kit (Plant) is designed specifically for genomic DNA isolation from plant samples. This unique buffer system ensures total DNA with high yield and good quality from samples. The spin column system is designed to purify or concentrate DNA products which have been previously isolated using buffers. The entire procedure can be completed in 1 hour without phenol/chloroform extraction. Purified DNA is suitable for use in PCR or other enzymatic reactions.

• Sample: up to 100 mg of fresh plant tissue up to 50 mg of dry plant tissue.
• Format: spin column.
• Column capacity: up to 50 µg.
• Operation time: within 60 minutes.

Kit Contents

• Delivering high-quality genomic DNA with the fast procedure.
• Ready-to-use genomic DNA for high performance in any downstream application.
• Highly purified and high yield genomic DNA can be extracted from various tissue samples.
• Optimized tissue lysis buffer for the efficient lysis.
• Designed to rapidly purify high-quality DNA using spin column format.

Step 1 Sample Preparation

1. Cut off 50 mg of the fresh plant tissue or 25 mg of the dry plant tissue.
2. Grind the sample under liquid nitrogen to a fine powder using a mortar and pestle.

Step 2 Lysis

1. Add 500 µl of the Buffer PL and 0.5 µl of the RNase A (50 mg/ml) to the sample in the mortar and grind the sample until it is completely dissolved.
2. Transfer the dissolved sample to a 1.5 ml microcentrifuge tube.
3. Incubate at 75°C for 30 minutes. (invert the tube every 10 minutes)
4. Centrifuge at 14-16,000 x g for 5 minutes.
5. Transfer the supernatant to a new 1.5 ml microcentrifuge tube. #Pre-heat the Buffer BE to 75°C for Step 5 DNA Elution.

Step 3 DNA Binding

1. Add the same volume of the isopropanol to the clear supernatant from the previous step and vortex immediately for 5 seconds (eg. add 350 µl Isopropanol to the 350 µl supernatant)
2. Place a Column PC in a 2 ml Collection Tube.
3. Transfer the mixture to the Column PC.
4. Centrifuge at 14,000 x g for 30 seconds.
5. Discard the flow-through and place the Column PC back in the same Collection Tube.

Step 4 Wash

1. Add 400 µl of the Buffer W1 into the Column PC.
2. Centrifuge at 14,000 x g for 30 seconds.
3. Discard the flow-through and place the Column PC back into the same Collection tube.
4. Add 600 µl of the Buffer W2 (Ethanol added) into the Column PC.
5. Centrifuge at 14,000 x g for 30 seconds.
6. Discard the flow-through and place the Column PC back into the same Collection tube.
7. Centrifuge at 14,000 x g again for 2 minutes to remove the residual Buffer W2.

Step 5 DNA Elution

1. Transfer the dried Column PC to a new 1.5 ml microcentrifuge tube.
2. Add 50-200 µl of the Pre-Heated Buffer BE or TE into the center of the column matrix.
3. Let stand at 75°C for 3 minutes.
4. Centrifuge for 2 minutes at 14,000 x g to elute the purified DNA.

Refer to the table below to troubleshoot problems that you may encounter when purifying the genomic DNA with the kit.

• Buffers B and W1 contain irritants. Wear gloves when handling these buffers.
• Add 60 ml of the ethanol to the Buffer W2 before use.
• During operation, always wear a lab coat, disposable gloves, and protective equipment.
• Check buffers before use for salt precipitation. Redissolve any precipitate by warming to 37°C.
• Research Use Only. Not intended for any animal or human therapeutic or diagnostic uses.