Team:Chiba/Calendar-Home/25 August 2008

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24 August 2008 <|> 26 August 2008

Contents

Laboratory work

Team:Input

The following glycerol Stock was made.

BBa_J22136(insert check済み)

プレートに生えたコロニーをpickして、試験管で2ml培養(LB-Amp,37°C,12h)

培養した試験管から180μl,LB-Ampを880μlとって、OD=0.495にした。

Liquid culture(OD=0.495) 0.67μl
60%Glycerol 0.33μl
20%Glycerol Stock 1.00ml

Team:Communication

(24/8)--->Digestion
  1. Double Digestion:BBa_J04500(2007)
  2. Double Digesiton:BBa_R0010(2007)
  3. Single Digestion:BBa_R0010(2007)
  4. BBa_R0010(2007)
Sample No. 1234
Sample DNA 5511
EcoRI 0.50.5--
SpeI 0.50.5--
Buffer 1 11--
Buffer 3 --1-
BSA 11--
dH2O 227.5-
TOTAL 10μl10μl10μl1μl
--->Gel Check
Chiba-0825-1.JPG
Sample No. 1~34
Sample DNA 101
Loading Dye 21
dH2O -4
TOTAL 12μl6μl
From right;
  1. Double Digestion:BBa_J04500(2007) -> OK
  2. Double Digesiton:BBa_R0010(2007) -> ?(low)
  3. Single Digestion:BBa_R0010(2007) -> ?(low)
  4. BBa_R0010(2007) -> OK(low)


--->Digestion (Double Digestion)

  1. BBa_C0178
  2. BBa_C0170
  3. BBa_J04500(2007)
Sample No. 123
Sample DNA 33ng/μl×60μl33ng/μl×60μl100ng/μl×20μl
PstI 222
XbaI 22-
SpeI --2
Buffer 2 --5
Buffer 3 1010-
BSA 10105
dH2O 161616
TOTAL 100μl100μl52μl

Team:Output

PCR

No. ①②③④⑤
DNA tamplate 11111
FW primer 5(Venus_YFP_fwd)5(LacZ_fwd)5(LacZ_fwd)5(rLUC_fwd)5(fLuc_fwd)
VR primer 5(VR)5(LacZ_rev)5(LacZ_rev)5(VR)5(VR)
dNTPmix 1010101010
Thermo pol buffer 1010101010
DNA pol(VENT) 11111
dH2O 6868686868
TOTAL 100μl100μl100μl100μl100μl

-->Gel Check


No. ①②③④⑤
DNA tamplate 11111
loading Dye 11111
dH2O 44444
TOTAL 6μl6μl6μl6μl6μl

Mini prep


Transformation

  • BBa_E2030
  • PUC19
  • pGFPUV
  • pGEX Venus YFP
  • Membren Venus YFP

PCR

No. ①
DNA tamplate 1
FW primer(Venus) 5
VR primer 5
dNTPmix 10
Thermo pol buffer 10
DNA pol(VENT) 1
dH2O 68
TOTAL 100μl
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