September/11 September 2009

From 2009.igem.org

1.colony check

  (ligation sample No.)  (no. of colonies)
            43                    +++
           3-1M                    ++

2.ligation check

UP   :  24A 24B 24C 24D 33A 33B 33C 1-23L ladder 29 2-18F 31C 32A
DOWN :  34A 34B 34C 2-6E 35A 35B 1-2M ladder 40A 40B 1-6I 39 1-14N

ligation OK! (but 35A , 39)

3.Min prep
Result of Nanodrop concentration check

(sample No.) (concentration)
   14-A           14.8
   14-B           16.4
   22-A           16.9
   22-B            8.8
   41-A           31.0
   41-B           22.0
   42-A           22.8
   42-B           17.7

4.digestion and ligation
digestion with restriction enzyme

K204045

Vector		Insert
No.31	12	1-8I	6
SpeI	1	XbaI	1
PstI	1	PstI	1
No.2 Buffer	2	No.2	2
dH₂O	4	dH₂O	10
total	20uL	total	20uL

K204046

Vector		Insert
No.32	12	CFP(1-8K)	6
SpeI	1	XbaI	1
PstI	1	PstI	1
No.2 Buffer	2	No.2	2
dH₂O	4	dH₂O	10
total	20uL	total	20uL

K204017

Vector		Insert
1-6I	8	No.9	12
SpeI	1	XbaI	1
PstI	1	PstI	1
No.2 Buffer	2	No.2	2
dH₂O	8	dH₂O	4
total	20uL	total	20uL

5.ligation check & gel purification

UP   : 35A 1-2M 39 1-14N 27 1-6I 28 2-18H radder
DOWN : 17  45  46

No.27 ligation OK!

ligation sample No.17 , 45 , 46
↓
gel purification
↓
ligation 16 degree O/N

6. min prep Result of Nanodrop concentration check

(sample No.) (concentration)
    24A            74.2
      B            52.4
    29             71.1
    33A            59.3
      B            41.8
    34A            53.2
      B            47.9
    35B            34.5
    40A            29.9
      B            36.6

7.for ligation check digestion sample

14A 14B 22A 22B 41A 41B 42A 42B 
24A 24B 29 33A 33B 34A 34B 35B 40A 40B

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