August/22 August 2009

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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

Vector		Insert
p(RhlR)([http://partsregistry.org/wiki/index.php?title=Part:BBa_I14017 1-18L])	6	RBS+RhlR+terminater([http://partsregistry.org/wiki/index.php?title=Part:BBa_I1466 1-23J])	6
EcoRI	1	EcoRI	1
XbaI	1	SpeI	1
No.2	2	No.2	2
dH₂O	12	dH₂O	12
total	20uL	total	20uL

K204012

Vector		Insert
terminater([http://partsregistry.org/wiki/index.php?title=Part:BBa_B0015 1-23L])	6	RBS+cinR(③-C)	18
EcoRI	1	EcoRI	1
XbaI	1	SpeI	1
No.2	2	No.2	2
dH₂O	10	dH₂O	8
total	20uL	total	20uL

K204013

Vector		Insert
RBS([http://partsregistry.org/wiki/index.php?title=Part:BBa_B0034 1-2M])	10	RhlI([http://partsregistry.org/wiki/index.php?title=Part:BBa_C0170 2-8G])	8
SpeI	1	XbaI	1
PstI	1	PstI	1
No.2 Buffer	2	No.2	2
dH₂O	6	dH₂O	8
total	20uL	total	20uL

K204014

Vector		Insert
RBS([http://partsregistry.org/wiki/index.php?title=Part:BBa_B0034 1-2M])	8	LasI([http://partsregistry.org/wiki/index.php?title=Part:BBa_C0078 1-14H])	4
SpeI	1	XbaI	1
PstI	1	PstI	1
No.2 Buffer	2	No.2	2
dH₂O	8	dH₂O	13
total	20uL	total	20uL

K204015

Vector		Insert
terminater([http://partsregistry.org/wiki/index.php?title=Part:BBa_B0015 1-23L])	6	RBS+cinI(⑥-A)	8
EcoRI	1	EcoRI	1
XbaI	1	SpeI	1
No.2	2	No.2	2
dH₂O	10	dH₂O	8
total	20uL	total	20uL

↓
37°C, overnight

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