Team:Newcastle/1 July 2010

From 2010.igem.org

(Difference between revisions)
(Procedures)
 
(14 intermediate revisions not shown)
Line 1: Line 1:
{{Team:Newcastle/mainbanner}}
{{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.
The aim of this experiment was to determine if ''Bacillus subtilis'' 168 is able to produce urease and degrade urea.
-
===Procedures===
+
==Procedures==
* Please refer to [[Team:Newcastle/Urease test| urease test]]
* Please refer to [[Team:Newcastle/Urease test| urease test]]
-
===Inference===
+
==Inference==
Christensen's Urea Agar was formulated to detect and differentiate urolytic and urea degrading microorganisms.
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.
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 18: Line 19:
For actual results, please see [[Team:Newcastle/2 July 2010#Results|02.07.10 lab notebook.]]
For actual results, please see [[Team:Newcastle/2 July 2010#Results|02.07.10 lab notebook.]]
-
==LacI BioBrick Construction==
+
=Competent ''E. coli'' Production=
-
 
+
-
===Aims===
+
-
*To use PCR to extract ''lacI'' (promoter, ribosome-binding site (RBS) & coding sequence (CDS)) from plasmid pMutin4 and ligate into vector pSB1AT3 in front of red fluorescent protein (RFP).
+
-
 
+
-
===Materials===
+
-
*Competent ''E. coli'' (DH5alpha strain)
+
-
*pMutin4 plasmid
+
-
*pSB1AT3 plasmid
+
-
 
+
-
===Protocol===
+
-
* [[TeamNewcastleTransformation of E. coli|Transformation of ''E. coli'']] DH5alpha with pMutin4 and pSB1AT3 separately.
+
-
 
+
-
===Inference===
+
-
*''Ecoli'' DH5alpha is used as a bioreactor to replicate our plasmids to create a large stock ready for extraction from cells.
+
-
 
+
-
 
+
-
==Competent ''E. coli'' Production==
+
-
 
+
-
===Aims===
+
-
*To make a stock of competent ''E. coli'' (DH5alpha strain) ready for transformation.
+
-
===Materials===
+
==Aims==
-
*''E. coli'' DH5alpha strain
+
*To make a stock of competent ''E. coli'' (DH5α strain) ready for transformation.
-
===Protocol===
+
==Materials and Protocol==
-
* Set up [[Team:Newcastle/Growing a liquid culture| liquid culture]] of ''E. coli'' DH5alpha
+
* Set up [[Team:Newcastle/Growing a liquid culture| liquid culture]] consisting of ''E. coli'' DH5α.
-
===Inference===
 
-
*Grow up a stock of ''E.coli'' DH5alpha strain ready to be made competent
 
{{Team:Newcastle/footer}}
{{Team:Newcastle/footer}}

Latest revision as of 14:11, 25 October 2010

iGEM Homepage Newcastle University BacillaFilla Homepage Image Map

Contents

Urease Test

Aims

The aim of this experiment was to determine if Bacillus subtilis 168 is able to produce urease and degrade urea.

Procedures

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.

  • 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


Newcastle University logo.png    Newcastle cbcb logo.pngNewcastle Biomedicine logo.gif    Team Newcastle CEG logo.gif
Newcastle iww logo.jpg  UNIPV Pavia Logo.gif  Newcastle BBSRC.gif    Newcastle Genevision logo.png Newcastle WelcomeTrust.jpg
FaceBook Icon