Sodium dodecyl sulphate (SDS) 1% lysis buffer

A 1% SDS lysis buffer is a standard solution used in molecular biology for lysing (breaking open) cells to release their contents. Here's how to mix it:

Materials:

  • Sodium dodecyl sulfate (SDS) powder
  • Deionized water (dH2O)
  • Sodium hydroxide (NaOH) pellets (optional, for adjusting pH)
  • Balance
  • Graduated cylinder
  • Stir plate with stir bar (or equivalent for mixing)
  • Disposable container

Steps:

  1. Calculate the amount of SDS needed:
    • Decide on the final volume of your lysis buffer (e.g., 100mL, 500mL).
    • To make a 1% (w/v) solution, you need 1 gram of SDS per 100 mL of buffer. So, for a 500 mL solution, you'd need 5 grams of SDS.
  2. Weigh out the SDS:
    • Using a balance, carefully weigh out the required amount of SDS powder.
  3. Dissolve SDS in dH2O:
    • Add the weighed SDS to a graduated cylinder containing a small volume of dH2O (e.g., 20-30mL for a 500mL final volume).
    • Stir the solution using a stir plate with a stir bar until the SDS dissolves completely. The solution may appear cloudy initially, but it should become clear with proper mixing.
  4. Adjust the volume:
    • Once the SDS is dissolved, add dH2O to the graduated cylinder to reach the final desired volume of your lysis buffer.
  5. Adjust pH (optional):
    • While not always necessary, some protocols may require adjusting the pH of the lysis buffer. The optimal pH will depend on your specific experiment.
    • If required, use a small amount of 1M NaOH solution (prepared beforehand by dissolving 4 grams of NaOH pellets in 100mL dH2O) to adjust the pH using a pH meter. Add the NaOH solution dropwise while monitoring the pH until you reach the desired value.
  6. Store the buffer:
    • Once prepared, aliquot the lysis buffer into smaller volumes and store them at 4°C for short-term use or -20°C for long-term storage.

Safety:

  • SDS is an irritant, so wear gloves and eye protection when handling it.
  • NaOH is corrosive, so handle it carefully and dispose of waste according to your institution's guidelines.

Additional Tips:

  • You can find commercially pre-made 10x or 20x SDS solutions, which can be diluted with dH2O to obtain the desired concentration.
  • Depending on the specific application, your lysis buffer may include other components besides SDS. Consult your experimental protocol for any additional ingredients.