NeoPrep mini

Description

Neo Biotech's miniprep kits (NB-60-0001) offer a fast and efficient way to extract pure plasmid DNA from Escherichia coli.
The process includes alkaline lysis of bacterial cells and DNA binding to silica under high salt conditions.
The resulting DNA is ideal for PCR, automated fluorescent sequencing, and various enzymatic applications.

Storage conditions and reagents preparation

All kit components can be stored at room temperature (20-25 °C) and are stable till the expiry date. Before use, add 1 mL of Buffer A1 to the RNase A vial and vortex. Transfer the resulting solution into the Buffer A1 bottle and mix thoroughly. Buffer A1 with RNase should be stored at 4 °C for frequent use and at -20 °C for infrequent use. Add 32 mL of 100% molecular biology grade ethanol to each bottle of buffer A4. Buffer A2 may form a precipitate upon storage. If necessary, dissolve the precipitate by warming the solution at 37 °C. Buffers A3 and AY contain guanidine hydrochloride. Wear gloves and goggles when using this kit.

System Components

• Buffer A1 - 15 mL

• Buffer A2 - 15 mL

• Buffer A3 - 20 mL

• Buffer AY - 30 mL

• Buffer A4 (concentrate) - 8 mL

• Buffer AE (does not contain EDTA) - 15 mL

• RNase A - 5 mg

• Neo Biotech Spin Columns - 50

• Collection Tubes (2 mL) - 50

Growing of bacterial cultures:

LB medium is recommended for bacterial cell cultivation, but rich media like 2xYT or TB can also be used.
Rich media promotes faster growth and early entry into the stationary phase compared to LB.
However, using rich media may lead to more dead or starving cells, resulting in partially degraded plasmid DNA and potential contamination with chromosomal DNA.
Overgrown cultures can contain excessive bacterial material, affecting lysis and precipitation steps.
To start, select a single colony from a selective plate and inoculate 1-5 mL of LB medium with the appropriate antibiotic.
Incubate for 12-16 hours at 37°C with vigorous shaking.