Team:Chiba/Calendar-Home/19 September 2008

From 2008.igem.org

18 September 2008 <|> 20 September 2008

Laboratory work

Team:Communication

Sample Name	[http://partsregistry.org/Part:BBa_J04500 J04500]	[http://partsregistry.org/Part:BBa_R0010 R0010]	[http://partsregistry.org/Part:BBa_C0161 C0161]	Cin
Sample DNA(μL)	30	30	90	20
Loading Dye(μL)	6	6	18	4
TOTAL(μL)	36	36	108	24

J04500' (Plac+RBS)

R0010 (Plac)

C0161 (RBS+Lux)

C0261 (Lux+LVA)

--->Gel Extract

(3 wells for 1 sample)

note:ADB buffer,
left at 37°C,for 6 hours

--->Zymo Clean

elution
- [http://partsregistry.org/Part:BBa_J04500 BBa_J04500]-->27μL
- [http://partsregistry.org/Part:BBa_C0161 BBa_C0161]-->11μL

--->Gel Check

--->Ligation

Sample Name	insert+vector	insert	vector
[http://partsregistry.org/Part:BBa_C0161 C0161](μL)	3	3	-
[http://partsregistry.org/Part:BBa_J04500 J04500](μL)	4.5	-	4.5
ligase(μL)	1	1	1
Buffer(invitrogen)(μL)	3	2	2
NFW(μL)	3.5	4	2.5
TOTAL(μL)	15	10	10

--->Transformed above ligation onto a LB-amp plate.

note
- competent cell:XL10GOLD
- DNA:1μL,SOC:120μL

--->(20/9)Mini prep

time-delay test
- conditions

strain
Senders:XL10GOLD
Receiver:JW1908
temparature

30°C and Room Temparature

Team:Output

colony PCR
insert:[http://partsregistry.org/Part:BBa_S03119 BBa_S03119](×16)(marker:Kan)
insert:[http://partsregistry.org/wiki/index.php?title=Part:BBa_K084004 K084004](×5)(marker:Kan)
insert:[http://partsregistry.org/wiki/index.php?title=Part:BBa_K084005 K084005](×7)(marker:Kan)
insert:[http://partsregistry.org/wiki/index.php?title=Part:BBa_K084006 K084006](×5)(marker:Kan)