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)
Resuspend IAA in ethanol at a ratio of approximately 11 uL ethanol to 2 mg IAA. Vortex until dissolved.
Add mixture to ELGA water to achieve a final IAA concentration of 1.75 mg/mL (10mM).
Sterilize with 0.22 µm filter.
Separate into 50 uL aliquots and store at -20°C.
Indole-3-acetamide (Auxin precursor, IAM)
Resuspend IAM in ethanol at a ratio of approximately 20 uL ethanol to 1 mg IAM.
Add mixture to ELGA water to achieve a final IAM concentration of 1.75 mg/mL (10mM).
Sterilize with 0.22 µm filter.
Separate into 50 uL aliquots and store at -20°C.
50mM O6-Benzylguanine (O6BG)
Dissolve 12 mg of O6BG in 1ml of DMSO.
Sterilize with 0.22 µm filter.
Separate into 50 uL aliquots and store at -20°C.
25mM Guanine
Dissolve 7.5 mg of guanine in 2 ml of 0.2N NaOH/Water. (Guanine can not be dissolved with DMSO/Water)
Sterilize with 0.22 µm filter.
Separate into 50 uL aliquots and store at -20°C.
50 μM AP1903/Rimiducid (1000x stock)
Dissolve 1 mg of AP1903 in 14.2 mL of .1 % DMSO/water.
Sterilize with 0.22 µm filter.
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:
Make 5 mM HEPES from the 1M HEPES stock: 45 µL of 1M HEPES + 9 mL sterile water
Add 1 mL of 10 mg/mL puro to the 9 mL of 5 mM HEPES
Sterilize with 0.22 μM filer
Split into 50 μL aliquots and store at -20°C