Team:Chiba/jk/β/week 5

From 2008.igem.org

(Difference between revisions)
(19 September, 2008)
m (20 September, 2008)
 
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===19 September, 2008===
===19 September, 2008===
-->(?/9)
-->(?/9)
 +
'''[[Team:Chiba/protocol/gelcheck|Gel Check]]'''
 +
<table width="315" border="2" cellpadding="0" cellspacing="0" bordercolor="#000000">
 +
<tr>
 +
<td width="257">Sample Name</td>
 +
<td>J04500</td><td>R0010</td><td>C0161</td><td>Cin</td>
 +
</tr>
 +
<tr>
 +
<td>Sample DNA</td>
 +
<td>30</td><td>30</td><td>90</td><td>?</td>
 +
</tr>
 +
<tr>
 +
<td>Loading Dye</td>
 +
<td>6</td><td>6</td><td>18</td><td>?</td>
 +
</tr>
 +
<tr>
 +
<td>TOTAL</td>
 +
<td>36</td><td>36</td><td>108</td><td>24</td>
 +
</tr>
 +
</table>
 +
--->'''[[Team:Chiba/protocol/ligation/gelex|Gel Extract]]'''<br>
 +
(各サンプル well3つずつ)<br>
 +
*note:ADB buffer添加後,37℃,6h
 +
:--->'''[[Team:Chiba/protocol/DNA Purification/zymocleam|Zymo Clean]]'''
 +
*elution
 +
**J0450-->27
 +
**C0161-->11μL
 +
--->'''[[Team:Chiba/protocol/gelcheck|Gel Check]]'''
 +
:--->'''[[Team:Chiba/protocol/ligation/ligation|Ligation]]'''
 +
<table width="315" border="2" cellpadding="0" cellspacing="0" bordercolor="#000000">
 +
<tr>
 +
<td width="257">Sample Name</td>
 +
<td>insert+vector</td><td>insert</td><td>vector</td>
 +
</tr>
 +
<tr>
 +
<td>C0161</td>
 +
<td>3</td><td>3</td><td>-</td>
 +
</tr>
 +
<tr>
 +
<td>J04500</td>
 +
<td>4.5</td><td>-</td><td>4.5</td>
 +
</tr>
 +
<td>ligase</td>
 +
<td>1</td><td>1</td><td>1</td>
 +
</tr>
 +
<tr>
 +
<td>Buffer(invitrogen)</td>
 +
<td>3</td><td>2</td><td>2</td>
 +
</tr>
 +
<tr>
 +
<td>NFW?</td>
 +
<td>3.5</td><td>4</td><td>2.5</td>
 +
</tr>
 +
<tr>
 +
<td>TOTAL</td>
 +
<td>15</td><td>10</td><td>10</td>
 +
</tr>
 +
</table>
 +
:--->'''[[Team:Chiba/protocol/transformation|Transformation]]''' above ligation onto a LB-amp plate.
 +
*note
 +
**competent cell:XL10GOLD
 +
**DNA:1μL,SOC:120μL
-
 
+
--->(20/9)'''[[Team:Chiba/protocol/DNA Purification/sigma|Mini prep]]
Line 68: Line 129:
===20 September, 2008===
===20 September, 2008===
 +
--->(19/9)'''[[Team:Chiba/protocol/DNA Purification/sigma|Mini prep]]
 +
 +
--->'''[[Team:Chiba/protocol/gelcheck|Gel Check]]'''
 +
<table width="315" border="2" cellpadding="0" cellspacing="0" bordercolor="#000000">
 +
<tr>
 +
<td width="257">Sample Name</td>
 +
<td>Lagation(J04500+C0161)</td>
 +
</tr>
 +
<tr>
 +
<td>Sample DNA</td>
 +
<td>1</td>
 +
</tr>
 +
<tr>
 +
<td>Loading Dye</td>
 +
<td>1</td>
 +
</tr>
 +
<tr>
 +
<td>H<sub>2</sub>O</td>
 +
<td>4</td>
 +
</tr>
 +
<tr>
 +
<td>TOTAL</td>
 +
<td>6</td>
 +
</tr>
 +
</table>
 +
:From left;
 +
::# -> OK
 +
::# -> OK
 +
::# -> OK
 +
::# -> OK
 +
 +
:--->'''[[Team:Chiba/protocol/digestion|Digestion Test]]'''
 +
<table width="500" border="2" cellpadding="0" cellspacing="0" bordercolor="#000000">
 +
<tr>
 +
<td width="257">Sample No.</td>
 +
<td>1-4</td><td>5-8</td>
 +
</tr>
 +
<tr>
 +
<td>Sample DNA</td>
 +
<td>3</td><td>3</td>
 +
</tr>
 +
<tr>
 +
<td>XbaⅠ</td>
 +
<td>0.25?</td><td>0.25</td>
 +
</tr>
 +
<tr>
 +
<td>SpeⅠ</td>
 +
<td>0.5?</td><td>0.5</td>
 +
</tr>
 +
<tr>
 +
<td>Buffer 2</td>
 +
<td>2</td><td>2</td>
 +
</tr>
 +
<tr>
 +
<td>BSA</td>
 +
<td>2?</td><td>2?</td>
 +
</tr>
 +
<tr>
 +
<td>dH<sub>2</sub>O</td>
 +
<td>?</td><td>12.5</td>
 +
</tr>
 +
<tr>
 +
<td>TOTAL</td>
 +
<td>20</td><td>20</td>
 +
</tr>
 +
</table>
 +
 +
 +
 +
:--->'''[[Team:Chiba/protocol/gelcheck|Gel Check]]'''
 +
:<table width="315" border="2" cellpadding="0" cellspacing="0" bordercolor="#000000">
 +
<tr>
 +
<td width="257">Sample No.</td>
 +
<td>1-4</td><td>5~8</td>
 +
</tr>
 +
<tr>
 +
<td>Sample DNA</td>
 +
<td>20</td><td>20</td>
 +
</tr>
 +
<tr>
 +
<td>Loading Dye</td>
 +
<td>4</td><td>4</td>
 +
</tr>
 +
<tr>
 +
<td>TOTAL</td>
 +
<td>24</td><td>24</td>
 +
</tr>
 +
</table>
 +
:From left;
 +
::# -> OK
 +
::# -> OK
 +
::# -> OK
 +
::# -> OK
 +
::# -> OK
 +
::# -> OK
 +
::# -> OK
 +
::# -> OK
 +
 +
--->(?/9)'''[[Team:Chiba/protocol/transformation|Transformation]]'''
 +
--->(22/9)'''phenotype check
===[[Team:Chiba/jk/β/week 6|>next week]]===
===[[Team:Chiba/jk/β/week 6|>next week]]===

Latest revision as of 13:20, 1 October 2008

>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 check

>next week

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