"

From 2013.igem.org

Revision as of 12:32, 31 July 2013 (view source) Batou (Talk | contribs) ← Older edit		Revision as of 14:47, 31 July 2013 (view source) Batou (Talk | contribs) Newer edit →
Line 12:		Line 12:
	<b>Carbenicillin:</b> Put 150 mg of Carbenicilline in 15 mL of distilled water (10 mg per mL).<br>		<b>Carbenicillin:</b> Put 150 mg of Carbenicilline in 15 mL of distilled water (10 mg per mL).<br>
-	Filter with a 0,22 µm millipore,divide in 2 mL tubes and store at -20 °C.<br>	+	Filter with a 0,22 µm millipore,divide in 2 mL tubes and store at -4 °C.<br>
	Use 1 µL of Carbenicillin for 1 mL of solution.<br><br>		Use 1 µL of Carbenicillin for 1 mL of solution.<br><br>
	<b>Kanamycin:</b> Put 750 mg of Kanamycin in 15 mL of distilled water (50 mg per mL).<br>		<b>Kanamycin:</b> Put 750 mg of Kanamycin in 15 mL of distilled water (50 mg per mL).<br>
-	Filter with a 0,22 µm millipore,divide in 2 mL tubes and store at -20 °C.<br>	+	Filter with a 0,22 µm millipore,divide in 2 mL tubes and store at -4 °C.<br>
	Use 1 µL of Kanamycin for 1 mL of solution.<br><br>		Use 1 µL of Kanamycin for 1 mL of solution.<br><br>
	<b>Chloramphenicol:</b> Put 525 mg of Kanamycin in 15 mL of ethanol (35 mg per mL).<br>		<b>Chloramphenicol:</b> Put 525 mg of Kanamycin in 15 mL of ethanol (35 mg per mL).<br>
-	Filter with a 0,22 µm millipore,divide in 2 mL tubes and store at -20 °C.<br>	+	Filter with a 0,22 µm millipore,divide in 2 mL tubes and store at -4 °C.<br>
	Use 25 µL of Chloramphenicol for 1 mL of solution.<br><br>		Use 25 µL of Chloramphenicol for 1 mL of solution.<br><br>
	Carbenicillin inhibit cell wall synthesis, kanamycin interfer with mRNA traduction and chloramphenicol inhibit ribosomial peptidyl transferase activity.		Carbenicillin inhibit cell wall synthesis, kanamycin interfer with mRNA traduction and chloramphenicol inhibit ribosomial peptidyl transferase activity.

---

Revision as of 14:47, 31 July 2013

Solutions, Media and Petri Dishes

Antibiotics

Carbenicillin: Put 150 mg of Carbenicilline in 15 mL of distilled water (10 mg per mL).
Filter with a 0,22 µm millipore,divide in 2 mL tubes and store at -4 °C.
Use 1 µL of Carbenicillin for 1 mL of solution.

Kanamycin: Put 750 mg of Kanamycin in 15 mL of distilled water (50 mg per mL).
Filter with a 0,22 µm millipore,divide in 2 mL tubes and store at -4 °C.
Use 1 µL of Kanamycin for 1 mL of solution.

Chloramphenicol: Put 525 mg of Kanamycin in 15 mL of ethanol (35 mg per mL).
Filter with a 0,22 µm millipore,divide in 2 mL tubes and store at -4 °C.
Use 25 µL of Chloramphenicol for 1 mL of solution.

Carbenicillin inhibit cell wall synthesis, kanamycin interfer with mRNA traduction and chloramphenicol inhibit ribosomial peptidyl transferase activity.

LB and LB Agar

Mix 20 g of LB Broth in 1L of distilled water in a bottle of 1L, then put in autoclave.
Mix 17,5 g of LB Agar and in 500 mL of distilled water in a bottle of 1L, then put in autoclave.

CaCl2 and CaCl2+glycerol

Tris HCl

Disolve 4,6g of Trisbase in 30 mL of distilled water.
Adjust pH with concentrated HCl(~4 mL) until pH=7,5.
Add distilled water until 50 mL then put in autoclave.
If the solution has a yellow coloration, do the preparation again with better Trisbase.
Tris HCl is used to store DNA.