TFP1

• 100 mM RbCl
• 50 mM MnCl2
• 30 mM potassium acetate
• 10 mM CaCl₂
• 15%glycerol

TFP2

• 100 mM MOPS
• 50 mM RbCl
• 75 mM CaCl₂
• 15%glycerol

SOC medium

• 20 g/L Tryptophane
• 5 g/L Yeast extract
• 0.5 g/L NaCl
• 250 mM KCl
• 1M MgCl₂
• 50% (w/v) sterile glucose
• Filter the solution through a 0.2 µm filter.

3T3 medium

• 500 mL D-MEM 1x
• 50 mL FBS 100%
• 5 mL GlutaMax 100x
• 5 mL Penicillin (10k units/mL) /Streptomyocin (10k µg/mL )

Jurkat medium

• 2 mL MEM 1x
• 20 mL FBS 100%
• 2 mL GlutaMax 100x
• 0.4 mL Penicillin (10k units/mL) /Streptomyocin (10k µg/mL )
• 200 mL RPMI

Annexin V binding buffer

• 10 mM HEPES
• 140 mM NaCl
• 2.5 mM
• 250 mM CaCl₂
• pH 7.4
• Filter the solution through a 0.2 µm filter.

LB-Broth

• Product name: Difco™ LB Broth, Miller (Lurica-Bertani)
• Company: BD
• Lot: 4339957
• CAS:

Agarose

• Product name: Agarose
• Company: Sigma-Aldrich
• Lot: #SLBK3392V
• CAS: 9012-36-6

Glycerol

• Product: Glycerol anhydrous BioChemica
• Company: AppliChem
• Lot:
• CAS:

Plasmid Purification

• Product: ZR Plasmid Miniprep ™-Classic
• Company: Zymo Research
• Lot: ZRC182892
• Catalog No: D4016

Changes to the protocol included an extra step to dry the column before elution and the use of water for elution instead of elution buffer. For elution of large plasmids we used prewarmed water.

Gel Extraction

• Product: ZymocleanTM Gel DNA Recovery Kit
• Company: Zymo Research
• Lot: ZRC183055
• Catalog No: D4002

Changes to the protocol included an extra step to dry the column before elution and the use of water for elution instead of elution buffer. For elution of large plasmids we used prewarmed water.

Lactate Detection Kit

• Product: L-Lactic Acid
• Company: Megazyme
• Lot:
• Catalog No:

Changes to the protocol included an extra step to dry the column before elution and the use of water for elution instead of elution buffer. For elution of large plasmids we used prewarmed water.

The detection of lactate using this kit relies on two enzymatic reactions. In the first, L-lactate and NAD&sup+; were converted into pyruvate and NADH+H&sup+; by L-lactate dehydrogenase. In the coupled reaction, pyruvate is used with D-glutamate by the D-glutamate-pyruvate transaminase to have the equilibrium of the first reaction in favour of the pyruvate. The concentration of NADH is finally measured as the optical density at 340 nm (OD340).