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Please Note: The content below is not a direct copy from the original Word document. It was generated by Gemini 3 Pro by combining information from multiple sections. It serves as a demonstration of how we can improve the SOPs using Docusaurus and Markdown formatting to make them more readable and accessible.

Wet Lab Protocols

Plasmids & Cloning​

Freezing Plasmids

When getting a new plasmid, freeze it as bacterial stocks in the -80 using NEBstable cells.

  • Write the number on the top AND side.
  • Underline the number (e.g., 609 vs 606) to avoid orientation errors.
  • Sequencing: Use Plasmidsaurus. Pickup box is on the 2nd floor (Meyer building).
  • Ordering: When ordering from Yoav, check the sequence and backbone carefully.

DNA Gels​

Gel Storage

A DNA gel can be kept in liquid for a few hours. For overnight storage, wrap in clingfilm and refrigerate to prevent Gel-Red diffusion.

  • Imaging: Wipe the tray with Kimwipes (not paper towels) to avoid debris. Move images from the shared Dropbox folder to your personal folder immediately.
  • Disposal: Clean the bottle with a brush after pouring leftovers.

Transformations​

Primer Tips
  • Check the SWAT pYM primer collection before ordering.
  • Move SWAT primers to screw-cap tubes.
  • If ordering >1000 NIS, notify Hanni.
  • PCR: If Kapa Hifi fails, try adding 1µl DMSO or use Q5.
  • Colonies: Always streak to singles. Wait 3 days for negative colonies to stop growing.
  • Controls: Always use positive and negative controls in PCR.

Strains & Media​

Strain Expiration
  • G418/Hygro: 6 months
  • NAT/Amp/Kan: 1 year
  • 5FOA/MTX: 3 months
  • W303 Strains: Cannot grow on OMM Amino Acid mix.
  • Freezing: Use printed stickers for the cap and side. Add a round sticker on the side as backup.

Western Blots​

Antibodies
  • Dilute antibodies in 2% BSA/PBS + phenol red.
  • Discard diluted antibodies when they turn yellow or after 1 year.
  • Primary Abs: Label with Serial #, Dilution, and Date.
  • Blocking: Do not use milk for streptavidin/anti-phospho blots (use Fish Serum).
  • Membranes: Can be left in blocking/primary Ab over the weekend (4°C), but NEVER in secondary Ab.
  • Transfer: For proteins >175kDa, use Wet Transfer (not semi-dry).