August/8 October 2009

From 2009.igem.org

1.sequence check
Today we conducted PCR in order to sequence the parts we have built over the summer. Using the chain-termination method and standard sequencing primers from the Parts Registry, we obtained chain-terminated clones of each sample and put them through an automatic DNA sequencer. The results will be picked up and analyzed tomorrow.

2.colony check

sample        no. of colony
 2-6O                +++
  11                   +++ 
  20                   ++
  21                   +++
 1-15L               +++
  1-15J              +++
   52                    20
   67                     2
   68                     0
   69                     0

3.ligation

sample 
 70 , 71 , 74 , 75

4.digation

K204059

Vector		Insert
1-8E	6	1-8K	1
SpeI	1	XbaI	1
PstI	1	PstI	1
No.2	2	No.2	2
dH₂O	8	dH₂O	13
total	20uL	total	20uL

K204072

Vector		Insert
2-8E	2	31	6
EcoRI	1	EcoRI	1
XbaI	1	SpeI	1
No.2	2	No.2	2
dH₂O	12	dH₂O	8
total	20uL	total	20uL

↓
37 degree , 3hr
gel cut & ligation

5.transformation

sample 
70 , 71 , 74 , 75 , X4 , X4

6.color intensity check

plac+color , ptet+color

7.sequence check

sample 
22 , 47 , 40 , 25 , 51 , 50 , 31 , 32

back to NOTES