Team:TMU-Tokyo/Notebook experiment/4th

From 2013.igem.org

(Difference between revisions)
Line 129: Line 129:
<Br>
<Br>
<Br>
<Br>
 +
<div style="text-align:center;">
 +
<Table cellpadding="5">
 +
<Tr>
 +
<Td>Fragment</Td><Td>Fragment 3 - Ap<Br>(control)</Td><Td>Fragment 3 - ApCm</Td>
 +
</Tr>
 +
<Tr>
 +
<Td>Primer</Td><Td>CmNR-Not<Br>141-1</Td><Td>CmNR-Not<Br>141-1</Td>
 +
</Tr>
 +
<Tr>
 +
<Td>Templete</Td><Td>(Cell suspention)</Td><Td>(Cell suspention)</Td>
 +
</Tr>
 +
</Table>
 +
</div>
 +
Result.<Br>
 +
<IMG Src="https://static.igem.org/mediawiki/2013/b/bc/TMU-Tokyo_0924_1.jpg"><Br>
 +
1-16: Fragment 3 - ApCm<Br>
 +
<Br>
 +
<Br>
 +
P1 transduction<Br>
 +
<Table><Tr><Td>Fragment 2 to Fragment 1</Td></Tr></Table>
 +
<Table><Tr><Td>Fragment 1 to Fragment 2</Td></Tr></Table>
 +
Result.<Br>
 +
<IMG Src="https://static.igem.org/mediawiki/2013/5/56/TMU-Tokyo_0924_2.jpg"><Br>
 +
<IMG Src="https://static.igem.org/mediawiki/2013/b/b4/TMU-Tokyo_0924_3.jpg"><Br>
 +
<Br>
 +
<Br>
 +
Electro Poration<Br>
 +
<Table><Tr><Td>Fragment 1, 2, 3 - Ap, 3 - Cm, 3 - ApCm (in pSBC3-Km) to <i>Escherichia coli<i> DH1</Td></Tr></Table>
 +
Result.<Br>
 +
<IMG Src="https://static.igem.org/mediawiki/2013/e/eb/TMU-Tokyo_0924_4.jpg"><Br>
 +
 +
 +
<Br>
 +
<Br>
 +
<b>September 25th</b><Br>
<b>September 25th</b><Br>
 +

Revision as of 23:16, 27 September 2013



Experiment > 4th week


September 22th
Mini prep following plasmid.
fimE-pBAD(in pSB1A3), pSB1C3-Km, pSB1A3-Km
Result.

1: pSB1C3-Km, 2-4: pSB1A3-Km, 5-6: fimE-pBAD


Check PCR.
Fragment(control)Fragment 3 - ApCm
Primer141-49
SaApL
141-49
SaApL
TempleteEscherichia coli
MG1655 red
(Cell suspention)
(Cell suspention)
Result.

1-9: Fragment 3 - ApCm

Check PCR.
Fragment(control)Fragment 3 - ApCmFragment 3 - ApCm
Primer2 sets of the others141-1
141-20
141-1
141-74
TempleteEscherichia coli
MG1655 red
(Cell suspention)
(Cell suspention)
Result.

1-7: 141-1 and 141-20, 8: control(141-1 and 141-20),
9-15: 141-1 and 141-74, 16: control(141-1 and 141-74),


Digestion following plasmid.
pSB1C3-Km-Blunt


P1 Preparation following fragment.
Fragment 1, 2


September 23th
PCR following fragment.
FragmentFragment 3 - ApCm
PrimermhpA-CmC6
SaApL
TempleteFragment 3 - Cm
Ap(SaApR-SaApL)
Result.

1: Fragment 3 - Cm, 2: Fragment 3 - ApCm,
3: pSB1C3-Km-Blunt


Check PCR.
FragmentFragment 3 - Ap
(control)
Fragment 3 - ApCm
PrimerCmNR-Not
SaApL
CmNR-Not
SaApL
Templete(Cell suspention)(Cell suspention)
Result.

1-31: Fragment 3 - ApCm


Electro Pration
Fragment 3 - Cm to Escherichia coli MG1655 red.


P1 Preparation
Fragment 3 - ApCm


September 24th
P1 transdunction
Fragment 1, 2
Result.



FragmentFragment 3 - Ap
(control)
Fragment 3 - ApCm
PrimerCmNR-Not
141-1
CmNR-Not
141-1
Templete(Cell suspention)(Cell suspention)
Result.

1-16: Fragment 3 - ApCm


P1 transduction
Fragment 2 to Fragment 1
Fragment 1 to Fragment 2
Result.




Electro Poration
Fragment 1, 2, 3 - Ap, 3 - Cm, 3 - ApCm (in pSBC3-Km) to Escherichia coli DH1
Result.



September 25th
September 26th
September 27th
September 28th


>>Go to next week Back to last week<< >>Go to next week