Genomic DNA Extraction Kit (Plant)
No:DMGPL100 & 300
For research use only
Sample: up to 100 mg of fresh plant tissue or 50 mg of dry plant tissue
Format: spin column
Operation time: within 60 minutes
Introduction
The BioDiamond Genomic DNA Extraction Kit (Plant) was designed specifically for genomic DNA isolation from Plant samples. This unique buffer system ensures total DNA with high yield and good quality from samples and the spin column system was 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.
Kit Contents
NOTE
★ Add ethanol (96–100%) to Buffer W2, shake before use (see bottle label for volume).
★ Check Buffers before use for salt precipitation. Redissolve any precipitate by warming to 37°C.
★ Buffers W1 contain irritants. Wear gloves when handling these buffers.
Quality Control
In accordance with FairBiotech’s ISO-certified Total Quality Management System, the quality of the BioDiamond Genomic DNA Extraction Kit (Plant) is tested on a lot-to-lot basis to ensure consistent product quality.
Additional requirements
RNase A (50 mg/ml), Isopropanol, absolute ethanol, mortar and pestle, microcentrifuge tubes.
Protocol
Step 1 Sample Preparation
- Cut off 50 mg of fresh plant tissue or 25 mg of dry plant tissue. Grind the sample under liquid nitrogen to a fine powder using a mortar and pestle.
- Add 500 μl of PL reagent and 0.5 μl of RNase A (50 mg/ml) to the sample in the mortar and grind the sample until it is completely dissolved.
- Transfer the dissolved sample to a 1.5 ml microcentrifuge tube. Incubate at 75ºC for 30 minutes (invert the tube every 10 minutes).
- Centrifuge at 14-16,000 x g for 5 minutes. Transfer the supernatant to a new 1.5 ml microcentrifuge tube.
Step 3 DNA Binding
- Add the same volume of Isopropanol to the clear supernatant from the previous step and vortex immediately for 5 seconds (eg. add 350 µl Isopropanol to 350 µl supernatant)
- Place a PC Column in a Collection Tube. Transfer the mixture to the PC Column.
- Centrifuge at 14,000 x g for 30 seconds. Discard the flow-through and place the PC Column back in the Collection Tube.
- Add 400 µl of W1 Buffer into the PC Column. Centrifuge at 14,000 x g for 30 seconds.
- Discard the flow-through and place the PC Column back into the same Collection tube.
- Add 600 µl of W2 Buffer (Ethanol added) into the PC Column. Centrifuge at 14,000 x g for 30 seconds.
- Discard the flow-through and place the PC Column back into the same Collection tube.
- Centrifuge at 14,000 x g again for 2 minutes to remove residual W2 Buffer.
- Transfer the dried PC Column to a new 1.5 ml microcentrifuge tube.
- Add 50-200 µl of Pre-Heated EL Buffer or TE into the center of the column matrix.
- Let stand at 75ºC for 3 minutes.
- Centrifuge for 2 minutes at 14,000 x g to elute the purified DNA.
Troubleshooting
dmgpl100___300.pdf | |
File Size: | 252 kb |
File Type: |