Team:Chiba/jk/β/week 5

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>送受信班

>Protocols

Contents

Week 5

>last week

14 September, 2008

(13/9)--->Mini prep


Transformation
Competent cells : XL10G
  • BBa_T9002[http://partsregistry.org/Part:BBa_T9002](2007)
  • Plac+RhlI+LVA
  • Plac+CinI+LVA
--->(15/9)

15 September, 2008

Transformation
competent cells : JW1908
  • BBa_S03623 [http://partsregistry.org/Part:BBa_S03623]
  • BBa_K084007 [http://partsregistry.org/Part:BBa_K084007]
  • BBa_K084008 [http://partsregistry.org/Part:BBa_K084008]
  • BBa_K084009 [http://partsregistry.org/Part:BBa_K084009]
  • BBa_T9002 [http://partsregistry.org/Part:BBa_T9002]

(15/9)-->T9002 check(AHL sensitivity and time response test)

16 September, 2008

(15/9)->

  1. strain
    Senders:JW1908
    Receiver:JW1908
  2. temparature
    37℃ and 30℃

17 September, 2008

  1. strain
    Senders:JW1908
    Receiver:JW1908
  2. temparature
    37℃ and 30℃
  1. strain
    Senders:XL10GOLD
    Receiver:JW1908
  2. temparature
    37℃ and 30℃

18 September, 2008

  1. strain
    Senders:XL10GOLD
    Receiver:JW1908
  2. temparature
    37℃ and 30℃

19 September, 2008

-->(?/9) Gel Check

Sample Name J04500R0010C0161Cin
Sample DNA 303090?
Loading Dye 6618?
TOTAL 363610824

--->Gel Extract
(各サンプル well３つずつ)

  • note:ADB buffer添加後,37℃,6h
--->Zymo Clean
  • elution
    • J0450-->27
    • C0161-->11μL

--->Gel Check

--->Ligation

</tr>

Sample Name insert+vectorinsertvector
C0161 33-
J04500 4.5-4.5
ligase 111
Buffer(invitrogen) 322
NFW? 3.542.5
TOTAL 151010
--->Transformation above ligation onto a LB-amp plate.
  • note
    • competent cell:XL10GOLD
    • DNA:1μL,SOC:120μL

--->(20/9)Mini prep


  1. strain
    Senders:XL10GOLD
    Receiver:JW1908
  2. temparature
    30℃ and Room Temparature

20 September, 2008

--->(19/9)Mini prep

--->Gel Check

Sample Name Lagation(J04500+C0161)
Sample DNA 1
Loading Dye 1
H2O 4
TOTAL 6
From left;
  1. 　-> OK
  2. 　-> OK
  3. 　-> OK
  4. 　-> OK
--->Digestion Test
Sample No. 1-45-8
Sample DNA 33
XbaⅠ 0.25?0.25
SpeⅠ 0.5?0.5
Buffer 2 22
BSA 2?2?
dH2O ?12.5
TOTAL 2020


--->Gel Check
Sample No. 1-45~8
Sample DNA 2020
Loading Dye 44
TOTAL 2424
From left;
  1. 　-> OK
  2. 　-> OK
  3. 　-> OK
  4. 　-> OK
  5. 　-> OK
  6. 　-> OK
  7. 　-> OK
  8. 　-> OK

--->(?/9)Transformation

--->(22/9)phenotype chack

>next week

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