Small molecule stocks for TC

Antibiotic concentrations

Reference: Selection antibiotics (ThermoFisher)

Name	Temperature	Stock conc	Common working conc.	293T conc. KL 2023.12.11
Blasticidin	-20°C	10 mg/mL (1kx)	1 - 20 µg/mL	10 µg/mL (1:1000) by 4-5 dpt
Geneticin (G-418)	-20°C	? mg/mL (1kx)	200 - 500 µg/mL	400 µg/mL (1:100) - no death observed
Hygromycin	-20°C	50 mg/mL (250x)	200 - 500 µg/mL	200 µg/mL (1:250) by 6-8 dpt
Puromycin	-20°C	1-10 mg/mL (1-10kx)	0.1 - 10 µg/mL	1 µg/mL (1:10,000) by 3-4 dpt
Zeocin	-20°C	100 mg/mL (100x)	50 - 400 µg/mL	1,000 µg/mL (1:100) >8 dpt

Other stock and working concentrations

Name	Temperature	Stock conc	Common working conc.
Doxycycline	-20°C	1 mg/mL (1kx)	1 ng/mL - 1 µg/mL
Grazoprevir (GZV)	-80°C	1 mM (1kx)	1 nM - 1 µM
Polybrene (Hexadimethrine bromide)	-20°C	5 mg/mL (1kx)	5 - 8 µg/mL

Indole-3-acetic acid (Auxin, IAA)

1. Resuspend IAA in ethanol at a ratio of approximately 11 uL ethanol to 2 mg IAA. Vortex until dissolved.

2. Add mixture to ELGA water to achieve a final IAA concentration of 1.75 mg/mL (10mM).

3. Sterilize with 0.22 µm filter.

4. Separate into 50 uL aliquots and store at -20°C.

Indole-3-acetamide (Auxin precursor, IAM)

1. Resuspend IAM in ethanol at a ratio of approximately 20 uL ethanol to 1 mg IAM.

2. Add mixture to ELGA water to achieve a final IAM concentration of 1.75 mg/mL (10mM).

3. Sterilize with 0.22 µm filter.

4. Separate into 50 uL aliquots and store at -20°C.

50mM O6-Benzylguanine (O6BG)

1. Dissolve 12 mg of O6BG in 1ml of DMSO.

2. Sterilize with 0.22 µm filter.

3. Separate into 50 uL aliquots and store at -20°C.

25mM Guanine

1. Dissolve 7.5 mg of guanine in 2 ml of 0.2N NaOH/Water. (Guanine can not be dissolved with DMSO/Water)

2. Sterilize with 0.22 µm filter.

3. Separate into 50 uL aliquots and store at -20°C.

50 μM AP1903/Rimiducid (1000x stock)

1. Dissolve 1 mg of AP1903 in 14.2 mL of .1 % DMSO/water.

2. Sterilize with 0.22 µm filter.

3. Separate into 50 uL aliquots and store at -20°C.

Puromycin, 1 mg / mL (1,000x stock)

We use dissolved 10 mg/mL puromycin (Invivogen, cat. ant-pr-1). It comes as 10 tubes x 1 ml @ 10 mg/mL dissolved in HEPES buffer (5 mM, pH 7.3).

To dissolve a single 1 mL @ 10 mg/mL tube:

1. Make 5 mM HEPES from the 1M HEPES stock: 45 µL of 1M HEPES + 9 mL sterile water

2. Add 1 mL of 10 mg/mL puro to the 9 mL of 5 mM HEPES

3. Sterilize with 0.22 μM filer

4. Split into 50 μL aliquots and store at -20°C