Team:Chiba/Calendar-Home/26 August 2008
From 2008.igem.org
25 August 2008 <|> 27 August 2008
Contents |
Laboratory work
Team:Input
no work
Team:Communication
- (8/25)--->Gel Check
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--->Gel Extract
- --->Zymo Clean
- eluted with following amount of elution buffer
- 5μl
- 5μl
- 27μl
- --->Gel Check
- --->SAP
Sample No. 3 Sample DNA(μL) 26 SAP(μL) 1 SAP Buffer(μL) 3 TOTAL(μL) 30
- --->Zymo Clean
- Sample No.3 -->9μl
- --->Gel Check
Sample No.3 1 Loading Dye(μL) 1 dH2O(μL) 4 TOTAL(μL) 6 - Sample No.3 -> OK ・・・30ng?
- --->Ligation
Sample No. (1) (2) (3) (4) (5) insert①C0178(μL) 1 - - 1 - insert②C0170(μL) - 1 - - 1 vector③J04500(μL) - - 2 2 2 ligase(μL) 1 1 1 1 1 Buffer(μL) 2 2 2 2 2 dH2O(μL) 6 6 5 4 4 TOTAL(μL) 10 10 10 10 10
- --->Transformation
- insert①[http://partsregistry.org/Part:BBa_C0178 BBa_C0178]
- insert②[http://partsregistry.org/Part:BBa_C0170 BBa_C0170]
- vector③[http://partsregistry.org/Part:BBa_J04500 BBa_J04500]
- insert①[http://partsregistry.org/Part:BBa_C0178 BBa_C0178]+vector③[http://partsregistry.org/Part:BBa_J04500 BBa_J04500]
- insert②[http://partsregistry.org/Part:BBa_C0170 BBa_C0170]+vector③[http://partsregistry.org/Part:BBa_J04500 BBa_J04500]
Team:Output
- [http://partsregistry.org/Part:BBa_F2620 BBa_F2620]
- [http://partsregistry.org/Part:BBa_F2620 BBa_F2620]
- [http://partsregistry.org/Part:BBa_J63001 BBa_J63001]
Sample No. ① ② ③ ④ ⑤ DNA tamplate 1 1 1 1 >1 Loading Dye 1 1 1 1 1 dH2O 4 4 4 4 4 TOTAL 6 6 6 6 6 - vector:[http://partsregistry.org/Part:BBa_F2620 BBa_F2620]
Sample No. [http://partsregistry.org/Part:BBa_F2620 BBa_F2620] DNA tamplate 100 SpeⅠ 2 PstⅠ 2 BSA(x10) 13 Buffer2 13 TOTAL 130 - insert:[http://partsregistry.org/Part:BBa_J52008 BBa_J52008],[http://partsregistry.org/Part:BBa_E2030 BBa_E2030]
Sample No. [http://partsregistry.org/Part:BBa_J52008 BBa_J52008] [http://partsregistry.org/Part:BBa_E2030 BBa_E2030] DNA tamplate 30 30 XbaⅠ 1 1 PstⅠ 1 1 DpnⅠ 1 1 Buffer3 4 4 BSA(x10) 4 4 TOTAL 40 40
-->37℃ 3hour
-->SAP
-->37℃ 30min
-->65℃ 15min
-->Zymo Clean- vector:[http://partsregistry.org/Part:BBa_F2620 BBa_F2620] insert:[http://partsregistry.org/Part:BBa_J52008 BBa_J52008]①
- vector:[http://partsregistry.org/Part:BBa_F2620 BBa_F2620] insert:[http://partsregistry.org/Part:BBa_E2030 BBa_E2030]②
- Negative control:[http://partsregistry.org/Part:BBa_F2620 BBa_F2620]③
Sample No. ① ② ③ vector 5 5 5 inser 10 7.5 0 Ligase Buffer 4 4 4 Ligase 1 1 1 dH2O 0 2.5 10 TOTAL 20 20 20 -->RT 2hour
-->Transformation(XL10G)
-->37℃ over night
-->colony PCR
-->Mini prep
- --->Zymo Clean