129 mM TNC Dissolve 19 g Tri Sodium Citrate Dihydrate (FW 294.1) in 500 ml ddH2O. Adjust pH to 7.0 with 4 N HCl. Autoclave or sterile filter. Label with contents, initials and date and store at RT. Use 40 ml in 400 ml 1x PBS (Final 12.9 mM).
2% TX100 To 500 ml 1x PBS add 10 ml TX100 and 1 ml 0.1% Crystal Violet. Label with contents, initials and date and store at RT. Use 1:1 with infected samples (Final 1% TX100)
0.1% Crystal Violet Dissolve 0.2 g crystal violet in 100 ml methanol, 20 ml glacial acetic acid and 80 ml ddH2O. Stir for 2 hours and filter. Store at RT.
dFBS Stock is stored at -20°C in TC freezer J (500 ml bottles). Thaw FBS overnight in refrigerator. Heat inactivate in 56°C water bath for 1 hour. Make 50 ml aliquots into sterile Falcon tubes. Label with contents, initials and date and store at -20°C in TC freezer K. Use 50 ml in 500 ml media (Final 10% dFBS).
1x PBS 10x stock is stored at 4°C in cold room (500 ml bottles). Pour 400 ml of 10x PBS into very clean container rinsed with deionized water. Add deionized water to 4 L. Aliquot 400 ml into each of 10 sterile bottles. Label and put autoclave tape on cap (be sure to loosen caps). Autoclave in long liquid cycle.
Pen/Strep 10,000 IU - 10,000 µg/ml stock is stored at -20°C in TC freezer J (100 ml bottles). Do not dilute, just aliquot 5 ml into each of 20 sterile vials. Label with contents, initials and date and store at -20°C in TC freezer K. Use 5 ml in 500 ml media (Final 100 IU - 100 µg/ml).
200 mM L-Glutamine 200 mM stock is stored at -20°C in TC freezer J (100 ml bottles). Heat at 37°C to completely dissolve. Do not dilute, aliquot 5 ml into each of 20 sterile vials. Label with contents, initials and date and store at -20°C in TC freezer K. Use 5 ml in 500 ml media (Final 2 mM) .
1 M HEPES 1 M stock is stored at 4°C in TC refrigerator (100 ml bottles). Do not dilute. Use 5 ml in 500 ml media (Final 10 mM).
10 U/µl rIL-2 1000 U/ml stock is stored at -20°C in TC freezer K (10 ml bottles). Do not dilute, aliquot 500 µl into each of 20 sterile cryovials. Label with contents, initials and date and store at -20°C in TC freezer K. Use 20 µl in 10 ml media (Final 20 U/ml).
0.2 mg/ml PHA Only use Murex PHA (Abbott). 2 mg stock is stored at 4°C in TC refrigerator. Add 10 ml of sterile IMDM and mix well. Aliquot 1 ml into each of 10 sterile cryovials. Label with contents, initials and date and store at -20°C in TC freezer K. Use 2.5 ml in 500 ml media (Final 1 µg/ml).
500 µg/ml Polybrene Stock is stored at 4°C in TC refrigerator. Dissolve 50 mg polybrene in 100 ml 1x PBS. Aliquot 5 ml into each of 20 sterile vials. Label with contents, initials and date and store at -20°C in TC freezer K. Use 5 ml in 500 ml media (Final 5 µg/ml)
0.2 mg/ml Puromycin Stock is stored at -20°C in TC freezer K. Dissolve 2 mg puromycin in 10 ml 1x PBS. Filter sterilize. Aliquot 500 µl into each of 20 labeled sterile tubes. Store at -20°C in TC freezer K. Use 10 µl in 10 ml media (Final 200 ng/ml).
50 mg/ml G418 Sulfate Geneticin (G418 Sulfate) stock stored at -20°C in TC freezer K (solution) or at RT in TC room (powder). Need 21 ml of 50 mg/ml solution. Aliquot 3 ml in each of 7 sterile vials. Label with contents, initials and date and store at -20°C in TC freezer. Use 3 ml in 500 ml media (Final 300 µg/ml).
NMS Stock is stored at -20°C in TC freezer K (10 ml bottles). Do not dilute, aliquot 500 µl into each of 20 sterile cryovials. Label with contents, initials and date and store at -20°C in TC freezer K.
10 mg/ml Ciprofloxacin Stock is stored in TC refrigerator (solution) or at RT on TC counter top (powder). Dissolve 100 mg powder in 10 ml ddH2O, sterile filter, and store solution at 4°C. Use 500 µl in 500 ml media (Final 10 µg/ml).

 

For specifics on appropriate handling and waste procedures please see the online chemical SOPs or our waste and spill notebook located in room 352.


Department of Microbiology
School of Medicine
University of Washington
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