From 2010.igem.org

Christensen's Urea Agar was formulated to detect and differentiate urolytic and urea degrading microorganisms. The addition of gelatine peptone, dextrose and reduced content of buffer supports a luxuriant growth at an early stage. Urea is the substrate and can be degraded by certain organisms, which results in ammonia building. The ammonia makes the media alkaline and therefore the indicator phenol red will change from orange color to pink-red color.

Negative control - No color change (Orange color)
Test 1 (Duplicate) - Pink-red color
Test 2 (Duplicate) - Pink-red color

For actual results, please see 02.07.10 lab notebook.

Competent E. coli Production

Aims

To make a stock of competent E. coli (DH5α strain) ready for transformation.

Materials and Protocol

Set up liquid culture consisting of E. coli DH5α.

@@ Line 1: / Line 1: @@
 {{Team:Newcastle/mainbanner}}
-==Urease Test==
-===Aims===
+=Urease Test=
+==Aims==
 The aim of this experiment was to determine if ''Bacillus subtilis'' 168 is able to produce urease and degrade urea.
-===Materials===
+==Procedures==
-*Pipettes
+* Please refer to [[Team:Newcastle/Urease test| urease test]]
-*Universal tubes
-*Streaking loop
-*Christensen's Urea Agar
-**Make up 1 liter of the agar mixture containing:
-*# Gelatine peptone 1.0g
-*# Potassium dihydrogen phosphate 2.0g
-*# Sodium chloride 5.0g
-*# Agar 20g
-**Top up to 1 liter and apply gentle heating to dissolve. Sterilize at 115°C for at least 20min.
-**Add the following to the molten base:
-*# D(+)-Glucose 1.0g
-*# Phenol red, 0.2% 6ml
-**Note: Ensure that the work was done using aseptic technique.
-**Transfer 10ml of the molten base to universal tube and allow the agar to set in a slanted position.
-**Store the harden agar in the fridge.
-===Protocol===
+==Inference==
-# Perform the experiment using aseptic technique.
-# Pick up single colony of ''B. subtilis'' 168 and streaking it onto the slanted agar tube.
-# Loosen up the cap to allow air exchange between the interior of the tube and the external environment.
-# Incubate the tubes overnight at 37°C.
-# Set up the tubes as indicated:
-##Negative control - Without ''B. subtilis'' 168
-##Test 1 (Duplicate) - Inoculated with ''B. subtilis'' 168
-##Test 2 (Duplicate) - Inoculated with ''B. subtilis'' 168
-===Inference===
 Christensen's Urea Agar was formulated to detect and differentiate urolytic and urea degrading microorganisms.
 The addition of gelatine peptone, dextrose and reduced content of buffer supports a luxuriant growth at an early stage. Urea is the substrate and can be degraded by certain organisms, which results in ammonia building. The ammonia makes the media alkaline and therefore the indicator phenol red will change from orange color to pink-red color.
@@ Line 41: / Line 17: @@
 *Test 2 (Duplicate) - Pink-red color
-For actual results, please see 02.07.10 lab results.
+For actual results, please see [[Team:Newcastle/2 July 2010#Results|02.07.10 lab notebook.]]
+=Competent ''E. coli'' Production=
+==Aims==
+*To make a stock of competent ''E. coli'' (DH5α strain) ready for transformation.
-==LacI BioBrick==
+==Materials and Protocol==
+* Set up [[Team:Newcastle/Growing a liquid culture| liquid culture]] consisting of ''E. coli'' DH5α.
-===Aims===
-To use PCR to extract the ''lacI'' from pMutin4 plasmid.
 {{Team:Newcastle/footer}}

Team:Newcastle/1 July 2010

From 2010.igem.org

Latest revision as of 14:11, 25 October 2010

Contents

Urease Test

Aims

Procedures

Inference

Competent E. coli Production

Aims

Materials and Protocol