Team:Chiba/jk/β/week 4

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Contents

Week 4

>last week

7 September, 2008

-->(6/9) Digestion Test

Sample DNA 1 : BBa_R0010(unknown)[http://partsregistry.org/Part:BBa_R0010]
Sample DNA 2 : BBa_R0010(100ng/μL)[http://partsregistry.org/Part:BBa_R0010]
Sample No 1,2
Sample DNA 12
PstⅠ 0.5
SpeⅠ 0.5
Buffer 2 2
(×10)BSA 2
dH2O 3
TOTAL 20


--->Gel Check
Sample leftright
Sample DNA 55
Loading Dye 11
TOTAL 66

Left side:BBa_R0010 Sample No.1
Right side:BBa_R0010 Sample No.2

-->(8/9)

8 September, 2008

-->(7/9)

Gel Extract


--->Zymo Clean
->15μL?

9 September, 2008

-->(6/9)PCR of I9030

--->Gel Check

--->Gel Extract

--->Zymo Clean

10 September, 2008

-->(10/9)

Colony Count
  1. LuxI
  2. LuxI with LVA
  3. Las
  4. Background(R0010)
  5. Background(C0261)

Pick colony

Colony-PCR

Colony PCR of 10 colonies from ligation plate(9/9-(2)) and 3 colonies from ligation plate(9/9-(3)) one from control plate(BBa_F2620[http://partsregistry.org/Part:BBa_F2620](2007)).
DNA Template 1
dNTP mix 5
Foward Primer 0.3
Reverse Primer 0.3
taq DNA polymerase 0.5
Thermopol Buffer 3
Nuclease free water 20
TOTAL 30μL

??

95℃,5min -> ( 95℃,1min -> 52℃,1min -> 72℃,1min )・・・25cycles -> 72℃,10min -> 6℃

??

-->Gel Check

--->(28/8)Miniprep

elute 50μL

11 September, 2008

12 September, 2008

13 September, 2008

Gel Check
  1. BBa_C0161[http://partsregistry.org/Part:BBa_Co161](2007)
  2. BBa_J04500[http://partsregistry.org/Part:BBa_J04500](2007)
  3. BBa_R0010[http://partsregistry.org/Part:BBa_R0010](2007)
Chiba-0913.JPG
Sample No 1~3
Sample DNA 1
Loading Dye 1
dH2O 4
TOTAL 6
From left;


-->Digestion

  1. BBa_C0161[http://partsregistry.org/Part:BBa_Co161](2007)
  2. BBa_J04500[http://partsregistry.org/Part:BBa_J04500](2007)
  3. BBa_R0010[http://partsregistry.org/Part:BBa_R0010](2007)


Sample No
Sample DNA 5511
EcoRⅠ 0.5o.5--
SpeⅠ 0.50.5--
Buffer 1 11--
Buffer 3 --1-
BSA 11--
dH2O 227.5-
TOTAL 1010101


>next week

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