Boston University 2006:making antibiotic stocks
From 2006.igem.org
(Difference between revisions)
(in progress - off to go verify and test it.) |
(→Method) |
||
| (3 intermediate revisions not shown) | |||
| Line 8: | Line 8: | ||
==Method== | ==Method== | ||
| - | + | {| border="1" cellpadding="4" style="text-align:center" | |
| + | |+ Common antibiotic stock & working concentrations | ||
| + | |- | ||
| + | !width="120"|Antibiotic | ||
| + | !width="50"|Abrv. | ||
| + | !width="75"|Working | ||
| + | !width="75"|Stock | ||
| + | !Notes | ||
| + | !Color Code | ||
| + | |- style="background:#ff9900;" | ||
| + | | Ampicillin || Amp || 100 ug/mL || 100 mg/mL || || orange | ||
| + | |- style="background:#66ff66;" | ||
| + | | Chloramphenicol || Chm || 30 ug/mL || 30 mg/mL || in 100% ethanol || green | ||
| + | |- style="background:#ff6666;" | ||
| + | | Kanamycin || Kan || 50 ug/mL || 50 mg/mL || || red | ||
| + | |- style="background:#ffff66;" | ||
| + | | Tetracycline HCL || Tet || 50 ug/mL || 50 mg/mL || in 100% ethanol || yellow | ||
| + | |} | ||
| - | # | + | '''These directions are not correct yet.''' Antibiotic stocks based on dH20 should to be filter-sterilized prior to use. |
| - | # | + | |
| + | # add less than 5mL of dH20 to the 5mL falcon tube(s). | ||
| + | # multiply the mg of antibiotic listed in the stock column by 5 and add it to the dH20 in the falcon tube. | ||
| + | # dilute the solution to 5mL. | ||
| + | # filter sterilize if not using an antibiotic that is dissolved in ethanol. | ||
| + | ## inst. on how to filter sterilize. | ||
| + | # using a P1000, pipette 5 1mL aliquots from the falcon tube into 5 microcentrifuge tubes; store at -20C. | ||
==Notes== | ==Notes== | ||
* Store all the aliquots at -20C. | * Store all the aliquots at -20C. | ||
* Tet is light-sensative, so be sure to prevent exposure (i.e. wrap it in tinfoil or keep it in a dark box) | * Tet is light-sensative, so be sure to prevent exposure (i.e. wrap it in tinfoil or keep it in a dark box) | ||
Latest revision as of 19:50, 13 September 2006
Use the following protocol to produce 5 1mL aliquots of stock solution of your desired antibiotic. Use a 1:1000 dilution to reach a working concentration from the stock (i.e. take 5uL from the Amp stock and add to 5mL of LB broth to produce 100ug/mL Amp + LB broth).
Materials
- Powder of given antibiotic
- dH20 and/or 100% ethanol
- 5mL Falcon tube(s)
- 5 1.5mL microcentrifuge tubes (per 5mL falcon tube)
Method
| Antibiotic | Abrv. | Working | Stock | Notes | Color Code |
|---|---|---|---|---|---|
| Ampicillin | Amp | 100 ug/mL | 100 mg/mL | orange | |
| Chloramphenicol | Chm | 30 ug/mL | 30 mg/mL | in 100% ethanol | green |
| Kanamycin | Kan | 50 ug/mL | 50 mg/mL | red | |
| Tetracycline HCL | Tet | 50 ug/mL | 50 mg/mL | in 100% ethanol | yellow |
These directions are not correct yet. Antibiotic stocks based on dH20 should to be filter-sterilized prior to use.
- add less than 5mL of dH20 to the 5mL falcon tube(s).
- multiply the mg of antibiotic listed in the stock column by 5 and add it to the dH20 in the falcon tube.
- dilute the solution to 5mL.
- filter sterilize if not using an antibiotic that is dissolved in ethanol.
- inst. on how to filter sterilize.
- using a P1000, pipette 5 1mL aliquots from the falcon tube into 5 microcentrifuge tubes; store at -20C.
Notes
- Store all the aliquots at -20C.
- Tet is light-sensative, so be sure to prevent exposure (i.e. wrap it in tinfoil or keep it in a dark box)
