August/22 August 2009

From 2009.igem.org

1.transformed check
Checked the cell cultures transformed and plated out yesterday (8/21) for colony formation and made an approximate count of the number of colonies.

(ligation sample No.) (no. of colonies)
       ⑤                  failure
       ⑦                    100~
       ⑧                    100~
⑤ try back (digestion and ligation)
⑦,⑧ ligation rapid check


2.Min prep
Result of Nanodrop concentration check

(sample No.) (concentration)
    [http://partsregistry.org/wiki/index.php?title=Part:BBa_I732101 1-13N]         30.3 ng/uL
    [http://partsregistry.org/wiki/index.php?title=Part:BBa_C0170 2-8G]          15.5
    [http://partsregistry.org/wiki/index.php?title=Part:BBa_I13521 2-6O]          24.4
                    26.0
    
    ③-C          15.8
    ④-A          25.4
    ⑥-A          16.2


3.transfromation

ligation sample ⑨ , ⑩ , ⑪


4.digestion and ligation

K204005

VectorInsert
p(RhlR)([http://partsregistry.org/wiki/index.php?title=Part:BBa_I14017 1-18L])6RBS+RhlR+terminater([http://partsregistry.org/wiki/index.php?title=Part:BBa_I1466 1-23J])6
EcoRI1EcoRI1
XbaI1SpeI1
No.22No.22
dH2O12dH2O12
total20uLtotal20uL


K204012

VectorInsert
terminater([http://partsregistry.org/wiki/index.php?title=Part:BBa_B0015 1-23L])6RBS+cinR(③-C)18
EcoRI1EcoRI1
XbaI1SpeI1
No.22No.22
dH2O10dH2O8
total20uLtotal20uL


K204013

VectorInsert
RBS([http://partsregistry.org/wiki/index.php?title=Part:BBa_B0034 1-2M])10RhlI([http://partsregistry.org/wiki/index.php?title=Part:BBa_C0170 2-8G])8
SpeI1XbaI1
PstI1PstI1
No.2 Buffer2No.22
dH2O6dH2O8
total20uLtotal20uL


K204014

VectorInsert
RBS([http://partsregistry.org/wiki/index.php?title=Part:BBa_B0034 1-2M])8LasI([http://partsregistry.org/wiki/index.php?title=Part:BBa_C0078 1-14H])4
SpeI1XbaI1
PstI1PstI1
No.2 Buffer2No.22
dH2O8dH2O13
total20uLtotal20uL


K204015

VectorInsert
terminater([http://partsregistry.org/wiki/index.php?title=Part:BBa_B0015 1-23L])6RBS+cinI(⑥-A)8
EcoRI1EcoRI1
XbaI1SpeI1
No.22No.22
dH2O10dH2O8
total20uLtotal20uL



37°C, overnight

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