Team:Newcastle/2 September 2010

From 2010.igem.org

(Difference between revisions)
(→Transformation of Bacillius subtilis 168 with pGFP-rrnB containing filamentous cell part)
(→Preparation for pH acclimatisation of Bacillus subtilis 168)
 
(4 intermediate revisions not shown)
Line 1: Line 1:
{{Team:Newcastle/mainbanner}}
{{Team:Newcastle/mainbanner}}
-
=pH changing for HEPES=
+
=Preparation for pH acclimatisation of ''Bacillus subtilis'' 168=
We prepared 100mM of HEPES (it is a dibasic compound) at pH 7.0 and sent it for autoclaving.
We prepared 100mM of HEPES (it is a dibasic compound) at pH 7.0 and sent it for autoclaving.
-
N.B. During autoclaving we loose some volume of the solution.
 
-
 
-
=''yneA''=
 
-
 
-
==Aims==
 
-
 
-
The aim of the experiment is to test if the filamentous cell part was cloned into pGFP-rrnB successfully.
 
-
 
-
==Materials and protocols==
 
-
 
-
#Please refer to: [[Team:Newcastle/Qiagen_Minipreps#Plasmid_extraction| Plasmid extraction]].
 
-
#Please refer to:[[Team:Newcastle/Restriction_digests|Restriction digest]]. We used Nhe1 and Spe1 to remove the ''yneA'' from pGFP-rrnB.
 
-
#Please refer to:[[Team:Newcastle/Gel_electrophoresis| Gel electrophoresis]] for running all the digested plasmid fragments.
 
-
 
-
==Results==
 
-
 
-
[[Image:03.09.10.png#file|400px]]
 
-
 
-
'''Figure 1''': Gel electrophoresis result for restriction digest of pGFPrrnB and ''yneA'' with Nhe1 and Spe1.
 
-
 
-
* '''Lane 1''': 1kb DNA ladder
 
-
* '''Lane 2''': Tube 1
 
-
* '''Lane 3''': Tube 2
 
-
* '''Lane 4''': Tube 3
 
-
* '''Lane 5''': Tube 4
 
-
* '''Lane 6''': Tube 5
 
-
* '''Lane 7''': Tube 6
 
-
* '''Lane 8''': Tube 7
 
-
* '''Lane 9''': Tube 8
 
-
* '''Lane 10''': Tube 9
 
-
* '''Lane 11''': Tube 10
 
-
* '''Lane 12''': Tube 11
 
-
* '''Lane 13''': Tube 12
 
-
* '''Lane 14''': 1kb DNA ladder
 
-
 
-
==Conclusion==
 
-
 
-
The results show that the digest works, there is a band at approximately 541bp corresponding to ''yneA'' and a band at approximately 8.4kbp corresponding to pGFPrrnb in lanes 2, 3, 4, 6, 7, 8, 9, 11 and 12.
 
-
 
-
 
{{Team:Newcastle/footer}}
{{Team:Newcastle/footer}}

Latest revision as of 01:53, 28 October 2010

iGEM Homepage Newcastle University BacillaFilla Homepage Image Map

Preparation for pH acclimatisation of Bacillus subtilis 168

We prepared 100mM of HEPES (it is a dibasic compound) at pH 7.0 and sent it for autoclaving.

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
Retrieved from "http://2010.igem.org/Team:Newcastle/2_September_2010"