IBI Plant Isolate DNA Extraction Kit - 100ml
- Sample Size: up to 1g fresh plant tissue or 0.5g dry plant tissue
- Operation Time: 60min or less
IBI Plant Isolate Kit provides a quick and easy 3 step CTAB and chloroform based method to isolate total DNA (including genomic, mitochondrial, and chloroplast DNA) from a variety of plant species (including algae and cyanobacteria). This unique reagent is able to lyse most common plant samples as well as plant samples with a high polysaccharide content. The extracted DNA is suitable for routine PCR Screening, Real-Time PCR, Southern Blotting, Mapping, and RFLP. Phenol extraction is not required and the entire procedure can be completed within 50 minutes.
IBI Plant Isolate is tested on a lot-to-lot basis. 50mg of fresh Arabidopsis leaves are initially ground in IBI Plant Isolate. A 15µl aliquot of extracted genomic DNA from a 100µl eluate is analyzed by electrophoresis on a 1% agarose gel.
Sample: Up to 1g of fresh or 0.5g of dry plant tissue
Format: CTAB & Chloroform DNA Precipitation Method – Scalable
Yield: dependent on sample size
How do you remove excess oil from the plant samples? 1. Cut off 100 mg of fresh plant tissue or 50 mg of dry plant tissue. 2. Freeze the sample with liquid nitrogen. 3. Grind the sample to a fine powder using a mortar and pestle. 4. Add 400 μl of GP1 Buffer or GPX1 Buffer and 5 μl of RNAse A (96-well genomic DNA kit plant) or 1 ml of IBI Plant Isolate and 0.5 μl of RNAse A (IBI Plant Isolate kit) to the sample in the mortar. 5. Continue grinding the sample until it is completely dissolved then transfer the sample lysate to a 1.5 ml micro-centrifuge tube. 6. Incubate the sample lysate at 65°C for 10-30 minutes then centrifuge at 14,000-16,000 x g for 5 minutes. 7. A layer of oil will float on the top; transfer only the “clear supernatant” to a clean 1.5 ml micro-centrifuge tube then continue the protocol.
After using the Plant Isolate product we determined the DNA concentration by both absorbance and fluorescence and got widely varying results. Why? DNA concentration will not be affected by RNA contamination when using fluorescence. RNA contamination will affect the results of DNA concentration when determined by absorbance. Increase the amount of RNAse A being used.