From 2010.igem.org
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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}}
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Revision as of 01:51, 28 October 2010
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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