Paul.xu: Created page with "Working concentrations and stock solutions. {| width="550" |- bgcolor="000000" | {| cellspacing="1" cellpadding="2px" width="100%" |- bgcolor="eeeeee" | rowspan="3" | | colsp..."

2012-07-22T15:23:44Z

Created page with "Working concentrations and stock solutions. {| width="550" |- bgcolor="000000" | {| cellspacing="1" cellpadding="2px" width="100%" |- bgcolor="eeeeee" | rowspan="3" | | colsp..."

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Working concentrations and stock solutions.

{| width="550"
|- bgcolor="000000"
|
{| cellspacing="1" cellpadding="2px" width="100%"
|- bgcolor="eeeeee"
| rowspan="3" |
| colspan="2" rowspan="2" | Stock solution
| align="CENTER" colspan="4" | Working concentration:
|- bgcolor="eeeeee"
| colspan="2" | Strict control
| colspan="2" | Weak control
|- bgcolor="eeeeee"
| mg/ml:
|
| µg/ml
| dilution
| µg/ml
| dilution
|- bgcolor="ffffff"
| Ampicillin
| 50 (H2O*)
| -20oC
| 20
| 0.4 µl/ml
| 60-125
| 1.2-2.5 µl/ml
|- bgcolor="ffffff"
| Carbenicillin
| 50 (H2O, 50%EtOH)
| -20oC
| 20
| 0.4 µl/ml
| 60-200
| 1.2-4 µl/ml
|- bgcolor="ffffff"
| Chloramphenicol
| 34 (EtOH, MetOH)
| -20oC
| 25
| 0.7 µl/ml
| 170
| 5 µl/ml
|- bgcolor="ffffff"
| D-cycloserine*
| 10 (0.1M NaP pH8.0)
| 4oC
| 200
| 20 µl/ml
|
|
|- bgcolor="ffffff"
| Gentamycin
| 10 (H2O)
| -20oC
| 15
| 1.5 µl/ml
|
|
|- bgcolor="ffffff"
| Hygromycin B
| 10 (H2O)
| -20oC
| 10
| 1 µl/ml
| 400
| 40 µl/ml
|- bgcolor="ffffff"
| Kanamicin
| 50 (H2O)
| 4oC- -20oC
| 10
| 0.2 µl/ml
| 50
| 1 µl/ml
|- bgcolor="ffffff"
| Kasugamycin
| 10 (H2O)
| -20oC
| 1000
| 100 µl/ml
|
|
|- bgcolor="ffffff"
| Nalidixic acid
| 5 (H2O,pH to 11 /NaOH)
| -20oC
| 15
| 3.0 µl/ml
|
|
|- bgcolor="ffffff"
| Neomycin
| 10 (H2O)
| -20oC
| 100
| 10 µl/ml
| 800
| 80 µl/ml
|- bgcolor="ffffff"
| Rifampicin**
| 34 (MetOH)
| -20oC
| 150
| 4.4 µl/ml
|
|
|- bgcolor="ffffff"
| Spectinomycin
| 10 (H2O)
| -20oC
| 100
| 10 µl/ml
|
|
|- bgcolor="ffffff"
| Streptomicin
| 50 (H2O)
| -20oC
| 10
| 0.2 µl/ml
| 50
| 1 µl/ml
|- bgcolor="ffffff"
| Tetracycline***
| 5 - 12.5 (EtOH, 70%EtOH)
| -20oC
| 10
| 2 µl/ml
| 50 100
| 10 µl/ml 20 µl/ml
|}

|}

Unstable solution, prepare just before use; ** - Light sensitive; *** - Light sensitive, Mg2 - inhibitor;

{| width="610"
|-
|
Preparation of stock solutions. Ampicillin.

{| width="313"
|- bgcolor="000000"
|
{| cellspacing="1" cellpadding="2px" width="100%"
|- bgcolor="eeeeee"
| align="center" |
| Conc.
| 1ml
| 5ml
| 10ml
|
|- bgcolor="ffffff"
| Ampicillin
| 50mg/ml
| 50mg
| 250mg
| 0.5g
|
|- bgcolor="ffffff"
| H2O
|
| 1ml
| 5ml
| 10ml
|
|}

|}

*filter sterilization; or preparation from the sterile water and the sterile powder;
*store at -20oC in small quantities (about 10 freezing-thawing cycles);
*instead of H2O it is possible to use 50% glycerol. The advantage - solution is liquid at -20oC.
*some laboratories use 96% (or 50%) EtOH for ampicillin solubilization. There are two advantages in this method: (i) solution is liquid at -20oC, (ii) the solution is self sterilizing. But the disadvantage it very serious: ethanol is a weak catalyst of b-lactam ring disruption. So, solution is unstable.

Carbenicillin.

{| width="313"
|- bgcolor="000000"
|
{| cellspacing="1" cellpadding="2px" width="100%"
|- bgcolor="eeeeee"
| align="center" |
| Conc.
| 1ml
| 5ml
| 10ml
|
|- bgcolor="ffffff"
| Carbenicillin
| 50mg/ml
| 50mg
| 250mg
| 0.5g
|
|- bgcolor="ffffff"
| H2O
|
| 1ml
| 5ml
| 10ml
|
|}

|}

*filter sterilization; or preparation from the sterile water and the sterile powder;
*store at -20oC in small quantities (about 10 freezing-thawing cycles);
*200 µg/ml for pETpfu.

Chloramphenicol.

{| width="345"
|- bgcolor="000000"
|
{| cellspacing="1" cellpadding="2px" width="100%"
|- bgcolor="eeeeee"
| align="center" |
| Conc.
| 1ml
| 5ml
| 10ml
|
|- bgcolor="ffffff"
| Chloramphenicol
| 34mg/ml
| 34mg
| 170mg
| 340mg
|
|}

|}

*store dry powder at 4oC, solution at -20oC (in EtOH);
*BL21(DE3)pLyzS - 34µg/ml -> 1µl for 1ml of medium.

GE antibiotic mix (50x).

{| width="427"
|- bgcolor="000000"
|
{| cellspacing="1" cellpadding="2px" width="100%"
|- bgcolor="eeeeee"
| align="center" |
| Conc.
| 40ml
|
|- bgcolor="ffffff"
| Cefotaxime 25µg/ml
| 25µg/ml
| 1mg
|
|- bgcolor="ffffff"
| Ceftriaxone 25µg/ml
| 25µg/ml
| 1mg
|
|- bgcolor="ffffff"
| KPO4(pH 6.0) 0.1M
|
| 40ml
|
|}

|}

*filter sterilization; or preparation from the sterile KPO4 and the sterile powder;
*store at -200C, in small quantities (about 10 freezing-thawing cycles).

Tetracycline.

{| width="301"
|- bgcolor="000000"
|
{| cellspacing="1" cellpadding="2px" width="100%"
|- bgcolor="eeeeee"
| align="center" |
| Conc.
| 1ml
| 5ml
| 10ml
|
|- bgcolor="ffffff"
| Tetracycline
| 5mg/ml
| 5mg
| 25mg
| 50mg
|
|- bgcolor="ffffff"
| EtOH
|
| 1ml
| 5ml
| 10ml
|
|}

|}

*solution in EtOH, store at -20oC.
*Mg- antagonist, for the selection it is necessary to use Mg-free medium.

|}

[[Category:Buffer_Recipe]]

Antibiotics - Revision history

Paul.xu: Created page with "Working concentrations and stock solutions. {| width="550" |- bgcolor="000000" | {| cellspacing="1" cellpadding="2px" width="100%" |- bgcolor="eeeeee" | rowspan="3" | | colsp..."