Team:HokkaidoU Japan/Notebook/August19

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{{Template:HokkaidoU_Japan}}<div class="linkbar"><div class="prev">[[Team:HokkaidoU_Japan/Notebook/August18|August 18]]</div>[[Team:HokkaidoU_Japan/Notebook|Notebook]]<div class="next">[[Team:HokkaidoU_Japan/Notebook/August23|August 23]]</div></div>
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=Ligation=
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Ligated Vector with Heat shock promotor and RBS
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{|border="1px" class="protocol"
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|-
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!Parts
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!By comparison to marker
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!size (bp)
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!(ng) required
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!(uL) used
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|-
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|Vector
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|5 ng/uL
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|2996 bp
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|10 ng
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|2 uL
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|-
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|Heat shock promotor
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|25 ng/uL
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|800 bp
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|8 ng
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|0.3 uL
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|-
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|RBS
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|1 ng/uL
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|50 bp
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|1 ng
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|1 uL
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|-
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|style="border-top:1px solid #996;" colspan="4"|'''Total'''
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|style="border-top:1px solid #996;"|'''3.3 uL'''
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|}
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Ligation and [[Team:HokkaidoU_Japan/Protocols|transformation]] follows same protocol as previously
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=Competency Check=
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# Transfered 1 uL of pUC119 DNA (0.35 ug/uL) into 0.5 mL tube and added 2.5 uL of ADW (0.1 ug/uL)
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# Took new 1.5 mL tube and added 999 uL of ADW (0.1 ng/uL) to it plus 1 uL of [1]
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# Took new 1.5 mL tube and added 99 uL of ADW to it plus 1 uL of [2]
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*Added pUC119 DNA (1 pg/uL) to each DH5alpha tube of 50ul and 30ul also varying the amount of pUC119 DNA:1 uL, 10 uL, 100 uL
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*Transformed
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=Ligation and [[Team:HokkaidoU_Japan/Protocols|Transformation]] between Vector，Transformation=
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Because there weren't any colonies the previous day we suspected that there might be something wrong with vectors<br>
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* Mixed 2 uL of pSB1C3, 2 uL of ligation solution and 0.4 uL of T4 ligase
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* Transformed

Latest revision as of 08:29, 27 October 2010

Notebook

Ligation

Ligated Vector with Heat shock promotor and RBS

Parts By comparison to marker size (bp) (ng) required (uL) used
Vector 5 ng/uL 2996 bp 10 ng 2 uL
Heat shock promotor 25 ng/uL 800 bp 8 ng 0.3 uL
RBS 1 ng/uL 50 bp 1 ng 1 uL
Total 3.3 uL

Ligation and transformation follows same protocol as previously

Competency Check

  1. Transfered 1 uL of pUC119 DNA (0.35 ug/uL) into 0.5 mL tube and added 2.5 uL of ADW (0.1 ug/uL)
  2. Took new 1.5 mL tube and added 999 uL of ADW (0.1 ng/uL) to it plus 1 uL of [1]
  3. Took new 1.5 mL tube and added 99 uL of ADW to it plus 1 uL of [2]
  • Added pUC119 DNA (1 pg/uL) to each DH5alpha tube of 50ul and 30ul also varying the amount of pUC119 DNA:1 uL, 10 uL, 100 uL
  • Transformed

Ligation and Transformation between Vector，Transformation

Because there weren't any colonies the previous day we suspected that there might be something wrong with vectors

  • Mixed 2 uL of pSB1C3, 2 uL of ligation solution and 0.4 uL of T4 ligase
  • Transformed
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