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| - | =Subtilin Immunity= | + | =Transformation of ligated ''yneA'' into pGFPrrnB and pSB1C3= |
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| - | ==Gel Electrophoresis and Gel Extraction==
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| - | ===Aims===
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| - | Following yesterday's Phusion PCR reactions, we plan to first perform gel electrophoresis of the amplified DNA sequences to check if they are of the correct sizes. If they appear to be successful we will then perform gel extraction, and finally nanodrop to record the level of DNA.
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| - | ===Materials and protocol===
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| - | Please refer to the:
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| - | *[[Team:Newcastle/Gel_electrophoresis| gel electrophoresis]],
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| - | *[[Team:Newcastle/Gel_extraction| gel extraction]] and
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| - | * [[TeamNewcastleNanoDrop_Spectrophotometer| NanoDrop spectrophotometer]] protocols.
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| - | ===Results===
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| - | Figure #. Gel electrophoresis of the amplified PCR products
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| - | Lane 1: 1 Kb ladder
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| - | Lane 2: pSB1C3 (for Subtilin Immunity)
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| - | Lane 3: pSB1C3 (for rocF)
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| - | Lane 4: pVeg (for Subtilin Immunity)
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| - | Lane 5: terminator (for Subtilin Immunity)
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| - | Lane 6: pSpacoid (for rocF)
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| - | Lane 7: terminator (for rocF)
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| - | Lane 8: 100 bp ladder
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| - | ===Discussion===
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| - | ===Conclusion===
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| - | ==PCR of pSB1C3 for rocF and Subtilin Immunity==
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| - | ===Aims===
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| - | =Gel Extraction=
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| - | ==Aims==
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| - | To purify the PCR products of digested ''yneA'', pGFPrrnB and pSB1C3 from [[Team:Newcastle/19_August_2010|yesterday]].
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| - | ==Materials and Protocol==
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| - | Please refer to [[Team:Newcastle/Gel_extraction|gel extraction]] and [[TeamNewcastleNanoDrop_Spectrophotometer|nanodrop]].
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| - | ==Results==
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| - | ==Discussion==
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| - | ==Conclusion==
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| - | =Miniprep of ''yneA'', pGFPrrnB and pSB1C3=
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| - | ==Aim==
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| - | To produce stocks of ''yneA'', pGFPrrnB and pSB1C3.
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| - | ==Materials and Protocol==
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| - | Please refer to [[Team:Newcastle/Qiagen_Minipreps|miniprep]] and [[TeamNewcastleNanoDrop_Spectrophotometer|nanodrop]].
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| - | ==Results==
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| - | ==Discussion==
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| - | ==Conclusion==
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| - | =Transformation of Ligated Products=
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| | ==Aims== | | ==Aims== |
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| - | To produce more colonies of the ligated products of ''yneA'' with pGFPrrnB and pSB1C3. | + | To produce more colonies of the plasmid pGFPrrnB and pSB1C3 containing ''yneA'' with pGFPrrnB and pSB1C3 from[[Team:Newcastle/19_August_2010|yesterday]]. |
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| | ==Materials and Protocol== | | ==Materials and Protocol== |
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| | Please refer to [[Team:Newcastle/Transformation_of_E._coli|transformation of E. coli]]. | | Please refer to [[Team:Newcastle/Transformation_of_E._coli|transformation of E. coli]]. |
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| - | =Ligation of ''yneA'' with Vectors=
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| - | ==Aims==
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| - | To ligate ''yneA'' with pGFPrrnB and ''yneA'' with pSB1C3 (A repeat of [[Team:Newcastle/19_August_2010|yesterday]].
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| - | ==Materials and Protocol==
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| - | Please refer to [[Team:Newcastle/Ligation|ligation]].
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| - | ==Results, Discussion and Conclusion==
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| - | Please refer to [[Team:Newcastle/23_August_2010|23.08.10]].
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| | {{Team:Newcastle/footer}} | | {{Team:Newcastle/footer}} |
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