FastPure Bacteria DNA Isolation Mini Kit

This kit is mainly intended for genomic DNA extraction from bacteria (Gram-positive or Gram-negative) of a variety of sources. The kit is based on silica gel column purification technology that eliminates the need for extraction using phenol/chloroform organic solvents or time-consuming alcohol precipitation step. With this kit, RNA, proteins, lipids and other inhibitory impurities can be removed at the greatest extent. The DNA obtained can be directly used in PCR, qPCR, enzyme digestion and Southern blot, etc. FastPure gDNA Mini Columns III 100 Buffer GA: Provide an environment for enzymolysis...

06/Notes 1. Please add a specified amount of absolute ethanol to Buffer PB and Buffer PW according to the label before first use. 2 Check if there is any precipitation in Buffer GA and Buffer GB before use. If precipitates have formed, they can be re-dissolved in a 37°C water bath and mixed well before use. 3. For Gram-positive bacteria samples, self-prepared Lysozyme is required for the treatment of bacteria. The Lysozyme should be used at a final concentration of 20 mg/ml. Lysozyme is not provided with this kit but can be purchased separately. 4. The amounts of input shouldn't exceed range...

1. Take 1 - 5 ml of bacteria culture medium (less than 1.0 * 109 bacteria), centrifuge it at 10,000 rpm (11,500 * g) for 1 min, then discard the culture medium. ▲ The quantity of bacteria can be measured with a spectrophotometer. 1 OD600 = 1.5 * 109 bacteria. 2. Add 230 gl of Buffer GA and vortex to thoroughly suspend the bacteria. 3. Add 20 gl of Proteinase K and vortex for mixing. 4. Add 250 gl of Buffer GB, mix by vortexing, and incubate in a 70°C water bath for 10 min. ▲ The addition of Buffer GB may produce white precipitates. Generally, the precipitates will disappear during heating at...

1. Add 180 pl of absolute ethanol, vortex to mix well (floccules may appear), and collect the liquid on the tube cap by brief centrifugation. 2. Transfer the above mixture to a FastPure gDNA Mini Columns III (already fitted in a Collection Tube). Centrifuge at 12,000 rpm (13,400 x g) for 1 min, and discard the filtrate. 3. Add 500 pl of Buffer PB (check whether absolute ethanol has been added before use) to the spin column, centrifuge at 12,000 rpm (13,400 x g) for 1 min, and discard the filtrate. 4. Add 600 pl of Buffer PW (check whether absolute ethanol has been added before use) to the spin...

Clogged FastPure gDNA Columns III Reduce the sample amount to less than 1.0 × 109 bacteria. 2. Undigested substances in digested solution If the solution resulting from sample digestion contains visible particulate matter, centrifuge the solution at 12,000 rpm (13,400 × g) for 3 min to remove the undigested substances. Determine the bacteria amount according to the bacterial culture situation. Some bacteria are lower in concentration after being cultured, so their usage amount can be increased appropriately. 2. Incomplete cell wall disruption Increase the Lysozyme amount or extend the enzymatic...

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