August/12 August 2009

From 2009.igem.org

1.before Min prep
Checked the cell cultures transformed and plated out yesterday (8/10) for colony formation and made an approximate count of the number of colonies.

(plate number)-(location on plate) (no. of colonies)
             1-14F                      50<
             1-14H                      50<
             1-23J                       100~
             1-12C                       100~
             2-18F                       10

inoculate to iquid medium(5mL + Amp 5uL or Kan 25uL)

2.Transformation
to get new plasmid

each 4uL(DNA) , 1-2M , 1-12C : 2uL
1-2M Amp
2-8O
1-21F
2-21F
1-12A
2-8I
1-12C
2-24G
1-16G

1-18L Kan
2-11N

'8/11sample  10uL
RBS+2-8M(LasR)
EpsE+Terminater
(after 65℃ , 10min)

3.LB plate

LB(500uL)+Amp(2.5mL)  *25
                  +Kan(0.5mL)  *25

4.Min prep
Result of Nanodrop concentration check

(sample No.) (concentration)
I179005 (1-15N)             19.0 ng/uL 
K091101 (2-6P)               27.0 
J23100 (1-18C)              21.5 
C0070 (1-12H)               48.0
B0015 (1-23L)                23.7
                                         19.4  
R0040 (1-6I)                    9.3
 
I15009 (1-20F)                 71.3
K118013 (2-18F)               19.2
C0078 (1-14H)                 49.9
I1466 (1-23J)                   21.3
R0071 (1-12C)                  10.7
C0077 (1-14F)                  32

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