Team:Newcastle/2 September 2010

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=Preparation for pH acclimatisation of ''Bacillus subtilis'' 168=
=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.
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=Double digest to confirm filamentous cell part was cloned successfully=
 
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==Aims==
 
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The aim of the experiment is to test if the filamentous cell part was cloned into pGFP-rrnB successfully.
 
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==Materials and protocols==
 
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#Please refer to: [[Team:Newcastle/Qiagen_Minipreps#Plasmid_extraction| Plasmid extraction]].
 
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#Please refer to:[[Team:Newcastle/Restriction_digests|Restriction digest]]. We used Nhe1 and Spe1 to remove the ''yneA'' from pGFP-rrnB.
 
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#Please refer to:[[Team:Newcastle/Gel_electrophoresis| Gel electrophoresis]] for running all the digested plasmid fragments.
 
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==Results==
 
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[[Image:03.09.10.png#file|400px]]
 
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'''Figure 1''': Gel electrophoresis result for restriction digest of pGFPrrnB and ''yneA'' with Nhe1 and Spe1.
 
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* '''Lane 1''': 1kb DNA ladder
 
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* '''Lane 2''': Tube 1
 
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* '''Lane 3''': Tube 2
 
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* '''Lane 4''': Tube 3
 
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* '''Lane 5''': Tube 4
 
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* '''Lane 6''': Tube 5
 
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* '''Lane 7''': Tube 6
 
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* '''Lane 8''': Tube 7
 
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* '''Lane 9''': Tube 8
 
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* '''Lane 10''': Tube 9
 
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* '''Lane 11''': Tube 10
 
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* '''Lane 12''': Tube 11
 
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* '''Lane 13''': Tube 12
 
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* '''Lane 14''': 1kb DNA ladder
 
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==Conclusion==
 
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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.
 
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{{Team:Newcastle/footer}}
{{Team:Newcastle/footer}}

Latest revision as of 01:53, 28 October 2010

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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.

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