Reagents and instruments for immunology, cell biology and molecular biology.
 
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NeoPrep mini

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.