TSS Buffer- GMES modified recipe

The price is for a 100ml pack.

Transformation and Storage Solution (TSS) is used for the one-step preparation of competent E. coli cells and to transform the cells without heat-shock

0.22µm filtration plus 5min 275/365nm UV treatment sterilized. Can be stored at room temperature. Suggest storage at 4°C / 39.2 ? refrigerator.

This buffer recipe is modified by GMES, to maximise the transformation rate.

Components in 100ml

3g PEG 8000 4g PEG 6000 3g PEG 4000 0.476g MgCl2 5 mL DMSO 0.1g Sorbitol 90µg Phospholipid membrane weakening catalyst (zwitterion) 10mg viscosity modifiers

Add SOC to 100 mL

Preparation

Inoculate 1mL of SOC medium with E. coli strain of choice in a 1.5mL PP tube (snap-cap) and culture overnight at 37°C with rotation.

Prepare 100mL of liquid medium LB in a sterile 250mL Erlenmeyer flask.

Seed the 1mL culture into the 250mL flask and incubate at 37°C with 225 rpm rotation until the OD600 reach 0.3 to 0.4 (0.3 = OD600 = 0.4) ( approximately 2-4h).

Once the proper optical density has been achieved, transfer the 100mL of culture into 2x 50mL PP centrifuge tubes and centrifuge under 3,000g for 10 min at 4°C.

Resuspend each tube in 5mL of ice-chilled TSS buffer with gentle vortexing.

Chill TSS-suspended cells on ice for 15 min. Prepare/label/pre-chill 50x 1.5mL PP tubes (snap-cap) during this time.

Distribute 100~200 µL of TSS suspended cells to each 1.5mL PP tube while ensuring the cells remain well mixed.

Cells can be used immediately or stored at -80°C.

Notice

Please kindly notice that the package appearance may change upon different manufacturing lots.

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