Home About Us News Products Technical Support Download Selling Network Recruitment Feedback Contact Us
Product class
DNA Markers£¨Ladders£©
Plasmid Isolation
PCR Product/Gel Purification
Genomic DNA Isolation
RNA Isolation
Protein Analysis
Electrophoresis Products
Molecular Biology Reagent
DNA/RNA Spin column
Lab Instruments
Technical Support You are here:Home >> Technical Support
Yeast Genomic DNA Fast Mini Kit£¨Spin column£©
Cat#GS3041   50 preps
The Yeast Genomic DNA Purification Kit provides a simple and rapid method for high quality genomic DNA purification from yeast. The kit uses the silica membrane technology for simple and fast isolation of Genomic DNA without phenol/chloroform. After treatment by Lyticase the unique binding buffer/ Proteinase K to rapidly lyse cells and inactivate cellualar nuclease, then DNA selectively adsorbs to silicified membrane in high salt solution. Cellular metabolite and proteins etc. are removed by serial of elution- centrifugation steps. Finally purified DNA from silica membrane is eluted by low salt elution buffer. About 10-15¦Ìg high quality genomic DNA can be purified from 1-3ml of culture. DNA purified using this method is ready for downstream applications such as PCR, Southern blotting, and restriction digestion.
Kit Content
50 preps
Buffer YB
20 ml
Binding Buffer CB
11 ml
Inhibitor Removing Buffer IR
25 ml
Washing Buffer WB
15 ml
Add 60ml ethanol before use.
Elution Buffer EB
15 ml
Proteinase K£¨20mg/ml£©
Spin-column AC
Collection Tube£¨2ml£©
Storage and Stability
Proteinase K must be stored at -20¡ãC. All other components can be stored at 22-25¡ãC.Under these conditions, performance of all components of the kit are guaranteed at least 12 months from the date of purchase. Under cool ambient conditions, a precipitate may form in the Buffer CB and Buffer IR, heat at 37¡ãC to dissolve the precipitate before use.
1. Dilute 15ml Buffer WB with 60ml absolute ethanol before use and mix thoroughly. Please mark it to avoid repeated add.
2. Buffer CB and IR may form precipitation due to low storage temperatures. If necessary, dissolve the precipitation at 37¡ãC and then cool to room temperature before use.
3. Once receiving the kit, pls store Proteinase K at -20¡æ.
4. 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 30 min.  
3. High purity - Multi-elution can ensure high-purified DNA. The typical ratio of OD260/OD280 is 1.7¡«1.9, and the average length is 30kb-50kb. Purified DNA can be applied for PCR£¬Southern-blot and digestions directly.
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(37¡æ) before use.
3. Prepare Isopropanol, ¦Â-Mercaptoethanol(14M).
4. Prepare Lyticase (10000U/ml)
5. Prepare Sorbitol buffer (1M sorbitol; 0.1M Na2EDTA; 14mM¦Â-Mercaptoethanol)
Note: Add 0.1% ¦Â-Mercaptoethanol(14M) to the sorbitol buffer at the final concentration of 14Mm before use.
6. Buffer CB and IR contain the stimulating compound; please wear latex gloves to avoid skin, eyes and cloth to be contaminated. If that, please use water or physiological saline washing.
Add 60 ml absolute ethanol to 15 ml Buffer WB before use.
1. Harvest 1-3ml (not exceed 3¡Á107 cells)yeast culture, centrifuge at 12,000 rpm for 30 seconds to pellet the cells. Discard the supernat carefully so as not to disturb or dislodge the cell pellet.
2. Add 600¦Ìl Sorbitol buffer to the cell pellet and gently pipet up and down until cells are suspended,
Add 200units of Lyticase (not included) (stock solution 10U/¦Ìl) and invert the tube 25 times to mix. Incubate at 37¡ãC for 30 -60min (optional: on shaker) to digest the cell walls. Invert the sample occasionally during the incubation.
3. Centrifuge at 13,000rpm for 1min, discard the supernatant, and resuspend pellet in 180¦Ìl Buffer YB.
4. Add 20¦Ìl Proteinase K (20mg/ml), vortex to mix thoroughly.
5. incubate at 55¡æ until solution become clear.
Optional RNase A treatment: If RNA-free genomic DNA is required, add of 10¦Ìl of RNase A (25mg/ml) after step 5, mix well and incubate at room temperature for 5-10 min..
6. Add 200¦Ìl Buffer CB£¬immediately vortex to mix thoroughly, incubate at 70¡æfor 10 min.
7. After cooling to room temperature , add 100¦Ìl isopropyl alcohol£¬then vortex to mix thoroughly£¬maybe appear the flocculated precipitate.
8. 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.
9. Add 500¦Ìl Buffer IR£¬centrifuge at 12,000 rpm for 30 sec, and discard flow-through.
10. Add 700¦Ìl Buffer WB£¨please check if ethanol added!£©£¬centrifuge at 12,000 rpm for 30 sec, and discard flow-through.
11. Add 500¦Ìl Buffer WB£¬centrifuge at 12,000 rpm for 30 sec, and discard flow-through.
12. Place Spin-column AC back to Collection Tube£¬centrifuge at 13,000 rpm for 2min to remove all the ethanol in the column.
13. Take the Spin-column AC out, then put it into a clean tube, add 100¦Ìl buffer EB (incubate at 65-70¡æ 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 ¦Ìl, or the elution efficiency and DNA yield will be affected.
14. Keep DNA at 2-8¡æ. For long-term storage, please store at -20¡æ.
Home About Us News Products Technical Support Download Selling Network Recruitment Feedback Contact Us
Guangzhou Geneshun Biotech Ltd ÔÁICP±¸08201538ºÅ
Tel£º020-23312160 Fax£º020-87056567