Team:Chiba/Calendar-Home/8 September 2008

From 2008.igem.org

(Difference between revisions)

Yoshimi (Talk | contribs)
(New page: >Home | Notebook 7 September 2008 <|> 9 September 2008 =...)
Newer edit →

Revision as of 07:38, 24 October 2008

>Home | Notebook

7 September 2008 <|> 9 September 2008

Laboratory work

Team:Input

no work

Team:Communication

-->(7/9)

Gel Extract

--->Zymo Clean

->15μL?

9 September, 2008

-->(6/9)PCR of I9030

--->Gel Check

--->Gel Extract

--->Zymo Clean

10 September, 2008

-->(10/9)

Colony Count

LuxI
LuxI with LVA
Las
Background(R0010)
Background(C0261)

Pick colony

Colony-PCR

Colony PCR of 10 colonies from ligation plate(9/9-(2)) and 3 colonies from ligation plate(9/9-(3)) one from control plate(BBa_F2620[http://partsregistry.org/Part:BBa_F2620](2007)).

DNA Template	1
dNTP mix	5
Foward Primer	0.3
Reverse Primer	0.3
taq DNA polymerase	0.5
Thermopol Buffer	3
Nuclease free water	20
TOTAL	30μL

??

95℃,5min -> ( 95℃,1min -> 52℃,1min -> 72℃,1min )･･･25cycles -> 72℃,10min -> 6℃

??

-->Gel Check

--->(28/8)Miniprep

elute 50μL

Team:Output

SAP

[http://partsregistry.org/Part:BBa_R0040 BBa_R0040](ptet)①
[http://partsregistry.org/Part:BBa_R0010 BBa_R0010](pLac)⑤

Sample No.	①	⑤
DNA(digested sample)	7	6
SAP/td>	2	2
SAP Buffer	3	3
TOTAL	30	30
TOTAL	30	30

-->37℃ 1hour

-->65℃ 30min

-->DNA purification

-->21μl 6μl

-->Gel check

Sample No.	each
DNA(digested sample)	1
Loading Dye	1
dH₂O	4
TOTAL	6

vector:[http://partsregistry.org/Part:BBa_R0040 BBa_R0040](ptet)① 75ng/μl
insert:[http://partsregistry.org/Part:BBa_S03158 BBa_S03158](RBS+RhlR)② 33.6ng/μl
insert:[http://partsregistry.org/Part:BBa_S03156 BBa_S03156](RBS+lasR)③ 33.6ng/μl
insert:[http://partsregistry.org/Part:BBa_S03160 Ba_S03160](RBS+cinR)④ 38.4ng/μl
vector:[http://partsregistry.org/Part:BBa_R0010 BBa_R0010](pLac)⑤ 100ng/μl
insert:[http://partsregistry.org/Part:BBa_J52008 BBa_J52008](RBS+rLuc)⑥ 33.6ng/μl

Sample No.	①&②	①&③	①&④	①	⑤&⑥	⑤
DNA(vector)	3	3	3	3	5	2.1
DNA(insert)	3	3	3	-	3.1	-
Ligase	1	1	1	1	1	1
Ligase Buffer(x5)	2	2	2	2	2	2
dH₂O	1	1	1	4	8.9	4
TOTAL	10	10	10	10	20	10