Sign InSign Up

Preparation of TSS Solution

Updated 5/25/2019 10:07 PM
Step-by-step
by SoundBio iGEM (created by JJ Wheeler)

Introduction

Preparation of TSS solution for competent cell prep. See https://openwetware.org/wiki/TSS and https://www.ncbi.nlm.nih.gov/pmc/articles/PMC286873/

Materials

  • Equipment
    • Scale
    • Eppendorf tubes (autoclaved)
    • Fisher tubes (1 non-sterile, 1 sterile)
    • 0.2 μm filter
    • 10-mL syringe
  • Reagents
    • LB media
    • PEG-3350 (Other PEG sizes are ok) (final concentration: 10% wt/vol)
    • DMSO (final concentration: 5% vol/vol)
    • 1M MgCl2 (final concentration: 50mM Mg++)

Procedure

For making 10mL TSS:

  1. Add about 8mL of LB to a 15-mL tube.
  1. Weight out 1g of PEG-3350.
  1. Add the PEG to the LB and vortex until it is dissolved.
  1. Add 0.3mL of 1M MgCl2 to the solution and vortex briefly.

This amount of MgCl2 is taken from the Openwetware recipe. The original paper says 50mM Mg++, but this wouldn't be 50mM. It would be 30mM. Weird?

  1. Add 0.5mL of DMSO to the solution and vortex.
  1. Bring the final volume to 10mL using LB.
  1. Filter sterilize with a 0.2 um filter and 10mL syringe

Comments

Benchling
Learn More
Contact
  • Contact Us
  • 680 Folsom St, 8th Floor
    San Francisco, CA 94107
Legal

Benchling is actively tested against the latest versions of Chrome, Firefox, Safari, and Edge. It doesn't look like your current browser is supported - for more information, click here.