August/9 October 2009

From 2009.igem.org

We collected sequence data from the automated sequencer and used sequence alignment software to check the sequences against those obtained from the Parts Registry. The results were as follows:

Senders:
LuxI - mismatched
LasI - OK
RhlI - OK
CinI - unconfirmed (DNA sample concentration too low?)

Receivers:
LuxR - OK
LasR - mismatched
RhlR - OK
CinR - unconfirmed (DNA sample concentration too low?)

The DNA for Cin signaling system will be transformed, amplified and purified to obtain at higher concentration and PCR sequencing repeated on them. As time is running out, for the final circuit we shall use parts sent from Chiba University to replace our faulty ones.


2.colony check

 sample   no . of colony
   32                 +++
   X4                  ~30
   X4                    7 
   70                     0
   71                     4
   74                      5
   75                      7


3.min prep

sample   conc.
  1-15J    98 ng/uL 
  1-15L    87
  43         169
  44          172
  52          114
  65         214
  67         148
  67         126

4.transfromation

saple
59 , 72 , 68 , 69 
chiba team parts ( 1 , 2 , 3 , 4 , 5 , 6 , 7 , 8 , 9)


5.digation

K204058

VectorInsert
1-8E41-8I10
SpeI1XbaI1
PstI1PstI1
No.2 2No.22
dH2O12dH2O6
total20uLtotal20uL


K204062

VectorInsert
1-8E9331
SpeI1XbaI1
PstI1PstI1
No.2 2No.22
dH2O7dH2O15
total20uLtotal20uL


K204072

VectorInsert
1-1D5642
SpeI1XbaI1
PstI1PstI1
No.2 2No.22
dH2O9dH2O12
total20uLtotal20uL


K204077

VectorInsert
1-6I7642
SpeI1XbaI1
PstI1PstI1
No.2 2No.22
dH2O7dH2O12
total20uLtotal20uL


K204078

VectorInsert
1-2M61-15N12
SpeI1XbaI1
PstI1PstI1
No.2 2No.22
dH2O10dH2O4
total20uLtotal20uL


37 degree , 2hr
gel cut & purification
ligation


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