July

From 2008.igem.org

(Difference between revisions)
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<p><h3>07-04-2008</h3><br />
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<p>'''07-04-2008'''<br />
   <br />
   <br />
   '''Assess Pool'''<br />
   '''Assess Pool'''<br />
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   '''Left border:''' 1.board, 1.line<br />
  '''Right border:''' 3.board, 2.line<br />
  '''Remainders:''' 3.board, 3.line<br />
   '''Left border:''' 1.board, 1.line<br />
  '''Right border:''' 3.board, 2.line<br />
  '''Remainders:''' 3.board, 3.line<br />
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   <h3>07-06-2008'''</h3><br />
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   '''07-06-2008'''<br />
   '''Assessment of cell-culture'''<br />
   '''Assessment of cell-culture'''<br />
   -(DYT-Medium; ER 2738-cells)<br />
   -(DYT-Medium; ER 2738-cells)<br />

Revision as of 20:17, 9 October 2008


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__july


07-04-2008

Assess Pool
We mixed all oligos together, except for the modified, the left and the right borders and the remainders:

Oligos modified with flourophor: 1. board:
r-3t8f
r-5t10e
r-5t12e

Oligos modified with NIP: 1. board:
r-7t6f
r-7t8f
r-7t10f
r-5t8f
r-3t10e
r-7t6e
3. board:
r7t22e

Left border: 1.board, 1.line
Right border: 3.board, 2.line
Remainders: 3.board, 3.line

07-06-2008
Assessment of cell-culture
-(DYT-Medium; ER 2738-cells)
50ml DYT-Medium
+50 TET (25ng/ml -> 1:1000)
+ pick ER 2738-cells (from -80 degree freezer)
-> shake at 37 degree over night

07-07-2008


 1) Thin down overnight culture to OD~0.1
1:10 -> 0.66 = OD 660

5000µl = 0.66
x = 0.1 -> x = 758µl ~ 760µl in 50ml

We mixed 760µl cell culture with 50ml DYT and shaked both at 37°C; Inoculation after 95min

<p>2) Inoculation of cell culture with M13mp18 phages

 ~ 50ml       cell culture
+ 5µl M13mp18 phages (from -80°C)
-> 4h shaking at 37°C

3) Phage precipitation
1) centrifuging of the phage-solution at 5000 x g for 20 min.
2) after decanting supernatant dessolve the pellet in 7ml (1/7 of the supernatant volume) PEG/NaCl
-> The precipitation occurs at 4°C over night

Freiburg08 FT3.png

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