| 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). |
