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Plant Genomic DNA Mini Kit(Spin column)
Cat#GS3052   100 preps
The Plant Genomic DNA Kit is used to quickly and efficiently isolate high quality genomic DNA from dry or fresh plant tissue. This kit is based on the efficient release of genomic DNA by using a special cell lysis buffer. A rapid separation of genomic DNA from proteins, polyphenols, polysaccharides and lipids by selective adsorption of DNA to the silica membrane becomes possible. The system provides a fast, simple and efficient way for purification, with the features of high yield, high purity and requiring no phenol and chloroform. DNA purified using this method is ready for downstream applications such as PCR, Southern blotting, and restriction digestion.
Kit Content

100 preps
RNase A(10mg/ml)
Buffer AP1
50 ml
Buffer AP2
20 ml
Buffer AP3
25 ml
Add 50ml ethanol before use.
Washing Buffer WB
25 ml
Add 100ml ethanol before use.
Elution Buffer EB
15 ml
Spin-column AC
Collection Tube2ml

Storage and Stability
RNase A store at -20C. All other components can be stored at 22-25C. Under these conditions, performance of all components of the kit are guaranteed at least 12 months from the date of purchase.
1) Dilute Buffer WB and Buffer AP3 with absolute ethanol before starting
2) Buffer AP1 and Buffer AP3 may form precipitation due to low storage temperature If necessary, dissolve the buffer by warming to 37C until become clear. Cool to room temperature before use.
3) Please ensure the bottles tightly capped when not in use, preventing reagents evaporating, oxidation and pH changing.
1. Safe-No need of harmful phenol and ethanol precipitation.
2. Simple and rapid- One preparation can be completed in 60 min.  
3. High purity - Multi-elution can ensure high-purified DNA. The typical ratio of OD260/OD280 is 1.71.9, isolated DNA ranges from 30Kb to 50Kb and can be directly used for most downstream
applications, including PCR, Southern-blot, Restriction digestion reactions, etc.
4. Stable, high-quality silica membrane and ideal buffer system ensure the reproducible results
Before staring
Please read this section before your experiment.
1.All the centrifugation steps can be performed at room temperature. Use a traditional Centrifugal machine that the rotational speed can reach 13,000rpm, such as Eppendorf 5415C and others.
2. Set water bath to the required temperature(65) before use.
3. Buffer AP3 includes the stimulating compound. Please wear latex gloves, avoiding skin, eyes and cloth to be contaminated. If that, please use water or physiological saline washing.
4. No EDTA in Buffer EB, which will have no influence on down-stream reactions. Also you can use water when eluting, but please ensure PH>7.5 and store at -20. If for long-term storage, dissolve DNA in TE10mM Tris-HCl, 1mM EDTA, PH 8.0. Because EDTA will affect the down-stream reactions, dilute the solution before using.
Dilute Buffer WB and Buffer AP3 with absolute ethanol before starting
1. Freeze up to 100mg fresh tissueor 20 mg dryin liquid nitrogen, grind frozen tissue to powder
using mortar and pestle.
2. Transfer the resulting powder into a new tube and add 400 l of Buffer AP1 and 4l RNase A(10mg/ml), vortex the mixture thoroughly.
Optional: if the sample is rich in polysaccharides add 2% PVP40,000 to Buffer AP1; if the sample is rich in Polyphenols add 0.2%- mercaptoethanol to Buffer AP1. or add both to Buffer AP1
3. Incubate at 65C for 10 min, Invert the sample occasionally during the incubation.
4. Add 130l Buffer AP2, mix thoroughly, incubate the sample on ice for 5 min, then Centrifuge at 14000rpm for 5-10min, carefully transfer the supernatant to a new tube.
5. Add 1.5 volume Buffer AP3, mix thoroughly.
6. Transfer the above step solution (including the flocculated precipitate) into a Spin-column AC (place the Spin-column AC to Collection Tube), then centrifuge at 13,000rpm for 30-60 sec, discard flow-through.
7. Add 700l Buffer WBplease check if ethanol added!centrifuge at 12,000 rpm for 30 sec, and discard flow-through.
8. Add 500l Buffer WBcentrifuge at 12,000 rpm for 30 sec, and discard flow-through.
9. Place Spin-column AC back to Collection Tubecentrifuge at 13,000 rpm for 2 min to remove all the ethanol in the column.
10. Take the Spin-column AC out, then put it into a clean tube, add 100l buffer EB (incubate at 65-70 water-bath), incubate for 3-5 min at RT, centrifuge at 12,000 rpm for 1 min.
Add the flow-through back in the Spin-column AC, let it stand for 3-5 min at RT. Centrifuge at 12,000 rpm for 1 min.
Note: Please reduce elution volume to increase the purified DNA concentration. But the elution volume not less than 50 l, or the elution efficiency and DNA yield will be affected.
11. Keep DNA at 2-8. For long-term storage, please store at -20.
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