From 2010.igem.org

Arginine is an amino acid that is positively charged. Therefore if B. subtilis 168 is able to take up arginine, it will cause a pH change in the media. This would result in an increase in pH.

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Figure 1: Arginine test using pH indicator stick to measure pH changes of the media.

Time (in minutes)	Control (1)	Control (2)	Control (3)	Test (1)	Test (2)
0	pH 7	pH 7	pH 7	pH 7	pH 7
30	pH 7	pH 7	pH 7	pH 7	pH 7
60	pH 7	pH 7	pH 7	pH 7	pH 7
90	pH 7	pH 7	pH 7	pH 7	pH 7
120	pH 7	pH 7	pH 7	pH 8	pH 8
150	pH 7	pH 7	pH 7	pH 8	pH 8
180	pH 7	pH 7	pH 7	pH 9	pH 9
210	pH 7	pH 7	pH 7	pH 9	pH 9
240	pH 7	pH 7	pH 7	pH 9	pH 9
270	pH 7	pH 7	pH 8	pH 9	pH 9
300	pH 7	pH 7	pH 8	pH 9	pH 9

Table 1: Arginine test using pH indicator stick to measure pH changes of the media. Table represents the change in pH over the time span of 300 minutes i.e. 5 hours.

Here,

Control (1) - LB media
Control (2) - LB media with 10 mM of arginine
Control (3) - LB media plus B. subtilis 168
Test (1) - LB media with 10 mM of arginine plus B. subtilis 168
Test (2) - LB media with 10 mM of arginine plus B. subtilis 168

Conclusion

B. subtilis 168 breaks down arginine to urea by producing arginase. The urea is then further broken down to ammonia and carbonate ions by urease. This will lead to an increase in pH. Both the test 1 and test 2, which contain B. subtilis 168 and 10 mM of arginine show an increase in pH from 7 to 9. While the control 1 and control 2, which contain no B. subtilis 168 remains at pH 7. The control 3 which contain B. subtilis 168 but without addition of arginine show an increase in pH from 7 to 8. This could be due to unidentified products that are secreted by the bacteria.

Therefore this experiment indicates that B. subtilis 168 is able to utilise arginine, and thus increase the overall pH of the media. The production of carbonate ion will lead to its binding with calcium ions provided in the media leading to formation of calcium carbonate.

@@ Line 1: / Line 1: @@
 {{Team:Newcastle/mainbanner}}
-===Aims of genomic DNA extraction experiment===
+=Arginine experiment=
-The aim of today's experiment is to extract genomic DNA from ''Bacillus subtilis'' strain 3610,genes from which will be needed for the swarming biobrick.
-====Procedure====
+==Aim==
-* Please refer to [[Team:Newcastle/DNA extraction| DNA extraction of ''Bacillus subtilis'']]
-====Discussion====
-At the end of the DNA rpecipitation step, we did not observe any small white pellet.
-====Conclusion====
-If the experiment has failed the experiment will be redone on Monday, 26th July, 2010.
-===Aim of Arginine experiment===
 The aim of this experiment is to determine whether ''B. subtilis'' 168 is able to take up external arginine.
-====Procedure====
+==Procedure==
 * Please refer to [[Team:Newcastle/Arginine test| Arginine test]]
-====Results====
+==Results==
-Arginine is an amino acid that is acidic. Therefore if ''B. subtilis'' 168 is able to take up arginine, it will cause a pH change in the media. This would result in an increase in pH.
+Arginine is an amino acid that is positively charged. Therefore if ''B. subtilis'' 168 is able to take up arginine, it will cause a pH change in the media. This would result in an increase in pH.
-|[[Image:Newcastle_arginine_test_230710.png|400px ]]
+|[[Image:Newcastle_arginine_test_230710.png|500px ]]
 '''Figure 1:''' Arginine test using pH indicator stick to measure pH changes of the media.
@@ Line 113: / Line 103: @@
 |}
-'''Table 1:''' Arginine test using pH indicator stick to measure pH changes of the media. Table represents the change in pH over the time span of 300 minutes i.e. 5 hours. Here,
+'''Table 1:''' Arginine test using pH indicator stick to measure pH changes of the media. Table represents the change in pH over the time span of 300 minutes i.e. 5 hours.
+Here,
 #Control (1) - LB media
 #Control (2) - LB media with 10 mM of arginine
@@ Line 120: / Line 112: @@
 #Test (2) - LB media with 10 mM of arginine plus ''B. subtilis'' 168
-====Conclusion====
+==Conclusion==
-''B. subtilis'' 168 breakdowns arginine to urea by producing arginase. The urea is then further broken down to ammonia and carbonate. This will lead to an increase in pH. Both the test 1 and test 2, which contain ''B. subtilis'' 168 and 10 mM of arginina show an increase in pH from 7 to 9. While the control 1 and control 2, which contain no ''B. subtilis'' 168 remains at pH 7. The control 3 which contain ''B. subtilis'' 168 but without addition of arginine show an increase in pH from 7 to 8. This could be due to unidentified products that are secreted by the bacteria.
+''B. subtilis'' 168 breaks down arginine to urea by producing arginase. The urea is then further broken down to ammonia and carbonate ions by urease. This will lead to an increase in pH. Both the test 1 and test 2, which contain ''B. subtilis'' 168 and 10 mM of arginine show an increase in pH from 7 to 9. While the control 1 and control 2, which contain no ''B. subtilis'' 168 remains at pH 7. The control 3 which contain ''B. subtilis'' 168 but without addition of arginine show an increase in pH from 7 to 8. This could be due to unidentified products that are secreted by the bacteria.
-Therefore this experiment have shown that ''B. subtilis'' 168 is able to utilise arginine, and thus increase the overall pH of the media. The overalll change in pH will cause calcium ions to precipitate with carbonate, forming calcium carbonate.
+Therefore this experiment indicates that ''B. subtilis'' 168 is able to utilise arginine, and thus increase the overall pH of the media. The production of carbonate ion will lead to its binding with calcium ions provided in the media leading to formation of calcium carbonate.
 {{Team:Newcastle/footer}}

Team:Newcastle/23 July 2010

From 2010.igem.org

Latest revision as of 21:00, 27 October 2010

Contents

Arginine experiment

Aim

Procedure

Results

Conclusion