IGEM:Harvard/2006/DNA nanostructures/Notebook/2006-8-15: Difference between revisions

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PEG:
PEG:
* GOAL: test 8% and 10% PEG precipitations
* GOAL: test 6%, 8%, 10% PEG precipitations
* incubated on ice for 15 min.
* incubated on ice for 15 min.
* spun at 16 k rcf at 4{{c}} for 10 min.
* spun at 16 k rcf at 4{{c}} for 10 min.
* carefully pipetted off supernatant
* carefully pipetted off supernatant
* resuspended "pellet" in 20 {{ul}} of water
* resuspended "pellet" in 50 {{ul}} of water




Line 15: Line 15:
| align="center" style="background:#f0f0f0;"|'''Trial'''
| align="center" style="background:#f0f0f0;"|'''Trial'''
| align="center" style="background:#f0f0f0;"|'''Final PEG %'''
| align="center" style="background:#f0f0f0;"|'''Final PEG %'''
| align="center" style="background:#f0f0f0;"|'''Gel'''
| align="center" style="background:#f0f0f0;"|'''Lanes'''
| align="center" style="background:#f0f0f0;"|'''Lanes'''
| align="center" style="background:#f0f0f0;"|'''20% PEG ({{ul}})'''
| align="center" style="background:#f0f0f0;"|'''20% PEG ({{ul}})'''
| align="center" style="background:#f0f0f0;"|'''5 M NaCl ({{ul}})'''
| align="center" style="background:#f0f0f0;"|'''5 M NaCl ({{ul}})'''
| align="center" style="background:#f0f0f0;"|'''Nanostructures ({{ul}})'''
| align="center" style="background:#f0f0f0;"|'''Nanostructures ({{ul}})'''
| align="center" style="background:#f0f0f0;"|'''water ({{ul}})'''
| align="center" style="background:#f0f0f0;"|'''Water ({{ul}})'''
| align="center" style="background:#f0f0f0;"|'''Total volume ({{ul}})'''
| align="center" style="background:#f0f0f0;"|'''Total volume ({{ul}})'''
|-
|-
| 1-0||0%||1||1||0||5||5||20||50
| 1-6||6%||1 (pellet)||15||5||20||10||50
|-
|-
| 1-2||2%||1||3||5||5||5||15||50
| 1-6||6%||2 (supernatant)||-||-||-||-||-
|-
|-
| 1-4||4%||1||5||10||5||5||10||50
| 1-8||8%||3 (pellet)||20||5||20||5||50
|-
|-
| 1-6||6%||1||7||15||5||5||5||50
| 1-8||8%||4 (supernatant)||-||-||-||-||-
|-
|-
| 1 kb+ ladder||||1||9||||||||||10
| 1-10||10%||5 (pellet)||25||5||20||0||50
|-
|-
| p7308||||1||10||||||||||10
| 1-10||10%||6 (supernatant)||-||-||-||-||-
|-
|-
| 1-8||8%||1||11||20||5||5||0||50
| 2-6||6%||7 (pellet)||15||5||20||10||50
|-
|-
| 2-0||0%||1||13||0||5||5||20||50
| 2-6||6%||8 (supernatant)||-||-||-||-||-
|-
|-
| 2-2||2%||1||15||5||5||5||15||50
| 2-8||8%||9 (pellet)||20||5||20||5||50
|-
|-
| 2-4||4%||1||17||10||5||5||10||50
| 2-8||8%||10 (supernatant)||-||-||-||-||-
|-
|-
| 2-6||6%||1||19||15||5||5||5||50
| 1kb+ ladder||-||11||-||-||-||-||10
|-
|-
| 2-8||8%||1||21||20||5||5||0||50
| p7308||-||12||-||-||-||-||10
|-
|-
| 3-0||0%||1||23||0||5||5||20||50
| 3-6||6%||11 (pellet)||15||5||20||10||50
|-
|-
| 3-2||2%||1||25||5||5||5||15||50
| 3-6||6%||12 (supernatant)||-||-||-||-||-
|-
|-
| 3-4||4%||1||27||10||5||5||10||50
| 3-8||8%||13 (pellet)||20||5||20||5||50
|-
|-
| 1 kb+ ladder||||1||29||||||||||10
| 3-8||8%||14 (supernatant)||-||-||-||-||-
|-
|-
| p7308||||1||30||||||||||10
| 3-10||10%||15 (pellet)||25||5||20||0||50
|-
|-
| 3-6||6%||1||31||15||5||5||5||50
| 3-10||10%||16 (supernatant)||-||-||-||-||-
|-
|-
| 3-8||8%||1||33||20||5||5||0||50
| 4-6||6%||17 (pellet)||15||5||20||10||50
|-
|-
| 4-0||0%||1||35||0||5||5||20||50
| 4-6||6%||18 (supernatant)||-||-||-||-||-
|-
|-
| 4-2||2%||1||37||5||5||5||15||50
| 4-8||8%||19 (pellet)||20||5||20||5||50
|-
|-
| 4-4||4%||1||39||10||5||5||10||50
| 4-8||8%||20 (supernatant)||-||-||-||-||-
|-
|-
| 4-6||6%||2||1||15||5||5||5||50
| 4-10||10%||1 (pellet)||25||5||20||0||50
|-
|-
| 4-8||8%||2||3||20||5||5||0||50
| 4-10||10%||2 (supernatant)||-||-||-||-||-
|-
|-
| 5-0||0%||2||5||0||5||5||20||50
| 5-6||6%||3 (pellet)||15||5||20||10||50
|-
|-
| 5-2||2%||2||7||5||5||5||15||50
| 5-6||6%||4 (supernatant)||-||-||-||-||-
|-
|-
| 1 kb+ ladder||||1||29||||||||||10
| 5-8||8%||5 (pellet)||20||5||20||5||50
|-
|-
| p7308||||1||30||||||||||10
| 5-8||8%||6 (supernatant)||-||-||-||-||-
|-
|-
| 5-4||4%||2||11||10||5||5||10||50
| 5-10||10%||7 (pellet)||25||5||20||0||50
|-
|-
| 5-6||6%||2||13||15||5||5||5||50
| 5-10||10%||8 (supernatant)||-||-||-||-||-
|-
|-
| 5-8||8%||2||15||20||5||5||0||50
| 6-6||6%||9 (pellet)||15||5||20||10||50
|-
|-
| 6-0||0%||2||17||0||5||5||20||50
| 6-6||6%||10 (supernatant)||-||-||-||-||-
|-
|-
| 6-2||2%||2||19||5||5||5||15||50
| 1kb+ ladder||-||11||-||-||-||-||10
|-
|-
| 6-4||4%||2||21||10||5||5||10||50
| p7308||-||12||-||-||-||-||10
|-
|-
| 6-6||6%||2||23||15||5||5||5||50
| 6-8||8%||13 (pellet)||20||5||20||5||50
|-
|-
| 6-8||8%||2||25||20||5||5||0||50
| 6-8||8%||14 (supernatant)||-||-||-||-||-
|-
|-
| 1 kb+ ladder||||1||29||||||||||10
| 6-10||10%||15 (pellet)||25||5||20||0||50
|-
|-
| p7308||||1||30||||||||||10
| 6-10||10%||16 (supernatant)||-||-||-||-||-
|}
|}

Revision as of 07:31, 15 August 2006

Mg2+, Oligo-Concentration Titration w/ c5.0 (cont'd)

3. PEG

PEG:

  • GOAL: test 6%, 8%, 10% PEG precipitations
  • incubated on ice for 15 min.
  • spun at 16 k rcf at 4[[:Category:{{{1}}}|{{{1}}}]] for 10 min.
  • carefully pipetted off supernatant
  • resuspended "pellet" in 50 μL of water


Trial Final PEG % Lanes 20% PEG (μL) 5 M NaCl (μL) Nanostructures (μL) Water (μL) Total volume (μL)
1-6 6% 1 (pellet) 15 5 20 10 50
1-6 6% 2 (supernatant) - - - - -
1-8 8% 3 (pellet) 20 5 20 5 50
1-8 8% 4 (supernatant) - - - - -
1-10 10% 5 (pellet) 25 5 20 0 50
1-10 10% 6 (supernatant) - - - - -
2-6 6% 7 (pellet) 15 5 20 10 50
2-6 6% 8 (supernatant) - - - - -
2-8 8% 9 (pellet) 20 5 20 5 50
2-8 8% 10 (supernatant) - - - - -
1kb+ ladder - 11 - - - - 10
p7308 - 12 - - - - 10
3-6 6% 11 (pellet) 15 5 20 10 50
3-6 6% 12 (supernatant) - - - - -
3-8 8% 13 (pellet) 20 5 20 5 50
3-8 8% 14 (supernatant) - - - - -
3-10 10% 15 (pellet) 25 5 20 0 50
3-10 10% 16 (supernatant) - - - - -
4-6 6% 17 (pellet) 15 5 20 10 50
4-6 6% 18 (supernatant) - - - - -
4-8 8% 19 (pellet) 20 5 20 5 50
4-8 8% 20 (supernatant) - - - - -
4-10 10% 1 (pellet) 25 5 20 0 50
4-10 10% 2 (supernatant) - - - - -
5-6 6% 3 (pellet) 15 5 20 10 50
5-6 6% 4 (supernatant) - - - - -
5-8 8% 5 (pellet) 20 5 20 5 50
5-8 8% 6 (supernatant) - - - - -
5-10 10% 7 (pellet) 25 5 20 0 50
5-10 10% 8 (supernatant) - - - - -
6-6 6% 9 (pellet) 15 5 20 10 50
6-6 6% 10 (supernatant) - - - - -
1kb+ ladder - 11 - - - - 10
p7308 - 12 - - - - 10
6-8 8% 13 (pellet) 20 5 20 5 50
6-8 8% 14 (supernatant) - - - - -
6-10 10% 15 (pellet) 25 5 20 0 50
6-10 10% 16 (supernatant) - - - - -
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