August/25 August 2009

From 2009.igem.org

(Difference between revisions)
(New page: 1.'''trandformation''' ligation sample No. 12 ,13 , 14 , 15 ↓ 65°C , 10min ↓ competent cell 100uL sample 10uL competent cell 100uL sample 2uL 2.'''...)
Line 16: Line 16:
Result of Nanodrop concentration check<br>
Result of Nanodrop concentration check<br>
  (sample No.) (concentration)
  (sample No.) (concentration)
-
     1-6I        13.5
+
     [http://partsregistry.org/wiki/index.php?title=Part:BBa_R0040 1-6I]         13.5
                     19.1
                     19.1
                     15.4
                     15.4
Line 22: Line 22:
                     18.9
                     18.9
        
        
-
     1-4E        15.0
+
     [http://partsregistry.org/wiki/index.php?title=Part:BBa_C0051 1-4E]         15.0
                     15.3
                     15.3
                     14.9
                     14.9
                    
                    
-
     2-16N        22.7
+
     [http://partsregistry.org/wiki/index.php?title=Part:BBa_K118006 2-16N]       22.7
                     17.7
                     17.7
                     23.1
                     23.1
          
          
-
     2-16H        13.8
+
     [http://partsregistry.org/wiki/index.php?title=Part:BBa_K118003 2-16H]       13.8
                     22.7
                     22.7
      
      
-
     2-16L        22.5
+
     [http://partsregistry.org/wiki/index.php?title=Part:BBa_K118005 2-16L]       22.5
                     31.5
                     31.5
Line 48: Line 48:
<table border="1" Frame="box">
<table border="1" Frame="box">
<tr><th colspan="2">Vector</th><th colspan="2">Insert</th></tr>
<tr><th colspan="2">Vector</th><th colspan="2">Insert</th></tr>
-
<tr><td>RBS(1-2M)</td><td>8</td><td>2-13N</td><td>6</td></tr>
+
<tr><td>RBS([http://partsregistry.org/wiki/index.php?title=Part:BBa_B0034 1-2M])</td><td>8</td><td>[http://partsregistry.org/wiki/index.php?title=Part:BBa_E2050 2-13N]</td><td>6</td></tr>
<tr><td>SpeI</td><td>1</td><td>XbaI</td><td>1</td><tr>
<tr><td>SpeI</td><td>1</td><td>XbaI</td><td>1</td><tr>
<tr><td>PstI</td><td>1</td><td>PstI</td><td>1</td><tr>
<tr><td>PstI</td><td>1</td><td>PstI</td><td>1</td><tr>
Line 60: Line 60:
<table border="1" Frame="box">
<table border="1" Frame="box">
<tr><th colspan="2">Vector</th><th colspan="2">Insert</th></tr>
<tr><th colspan="2">Vector</th><th colspan="2">Insert</th></tr>
-
<tr><td>RBS(1-2M)</td><td>8</td><td>YFP(1-8I)</td><td>6</td></tr>
+
<tr><td>RBS([http://partsregistry.org/wiki/index.php?title=Part:BBa_B0034 1-2M])</td><td>8</td><td>YFP([http://partsregistry.org/wiki/index.php?title=Part:BBa_E0430 1-8I])</td><td>6</td></tr>
<tr><td>SpeI</td><td>1</td><td>XbaI</td><td>1</td><tr>
<tr><td>SpeI</td><td>1</td><td>XbaI</td><td>1</td><tr>
<tr><td>PstI</td><td>1</td><td>PstI</td><td>1</td><tr>
<tr><td>PstI</td><td>1</td><td>PstI</td><td>1</td><tr>
Line 72: Line 72:
<table border="1" Frame="box">
<table border="1" Frame="box">
<tr><th colspan="2">Vector</th><th colspan="2">Insert</th></tr>
<tr><th colspan="2">Vector</th><th colspan="2">Insert</th></tr>
-
<tr><td>RBS(1-2M)</td><td>8</td><td>CFP(1-8K)</td><td>6</td></tr>
+
<tr><td>RBS([http://partsregistry.org/wiki/index.php?title=Part:BBa_B0034 1-2M])</td><td>8</td><td>CFP([http://partsregistry.org/wiki/index.php?title=Part:BBa_E0420 1-8K])</td><td>6</td></tr>
<tr><td>SpeI</td><td>1</td><td>XbaI</td><td>1</td><tr>
<tr><td>SpeI</td><td>1</td><td>XbaI</td><td>1</td><tr>
<tr><td>PstI</td><td>1</td><td>PstI</td><td>1</td><tr>
<tr><td>PstI</td><td>1</td><td>PstI</td><td>1</td><tr>

Revision as of 08:21, 29 September 2009

1.trandformation 

ligation sample No. 12 ,13 , 14 , 15 
↓
65°C , 10min
↓

competent cell 100uL
     sample     10uL 

competent cell 100uL
     sample      2uL


2.Min prep
Result of Nanodrop concentration check

(sample No.) (concentration)
    [http://partsregistry.org/wiki/index.php?title=Part:BBa_R0040 1-6I]         13.5
                    19.1
                    15.4
                    13.4
                    18.9
     
    [http://partsregistry.org/wiki/index.php?title=Part:BBa_C0051 1-4E]         15.0
                    15.3
                    14.9
                 
    [http://partsregistry.org/wiki/index.php?title=Part:BBa_K118006 2-16N]        22.7
                    17.7
                    23.1
       
    [http://partsregistry.org/wiki/index.php?title=Part:BBa_K118003 2-16H]        13.8
                    22.7
    
    [http://partsregistry.org/wiki/index.php?title=Part:BBa_K118005 2-16L]        22.5
                    31.5


3.ligation check

sample No. 9 , 10


4.digestion
digestion with restriction enzyme

K204019

VectorInsert
RBS([http://partsregistry.org/wiki/index.php?title=Part:BBa_B0034 1-2M])8[http://partsregistry.org/wiki/index.php?title=Part:BBa_E2050 2-13N]6
SpeI1XbaI1
PstI1PstI1
No.2 Buffer2No.22
dH2O8dH2O10
total20uLtotal20uL


K204020

VectorInsert
RBS([http://partsregistry.org/wiki/index.php?title=Part:BBa_B0034 1-2M])8YFP([http://partsregistry.org/wiki/index.php?title=Part:BBa_E0430 1-8I])6
SpeI1XbaI1
PstI1PstI1
No.2 Buffer2No.22
dH2O8dH2O10
total20uLtotal20uL


K204021

VectorInsert
RBS([http://partsregistry.org/wiki/index.php?title=Part:BBa_B0034 1-2M])8CFP([http://partsregistry.org/wiki/index.php?title=Part:BBa_E0420 1-8K])6
SpeI1XbaI1
PstI1PstI1
No.2 Buffer2No.22
dH2O8dH2O10
total20uLtotal20uL

↓
37°C, over night

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