IGEM:Harvard/2006/DNA nanostructures/Notebook/2006-7-24: Difference between revisions
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| 12||1.0||-||-||10.0** (3.2.Lb) ||2.0||2 | | 12||1.0||-||-||10.0** (3.2.Lb) ||2.0||2 | ||
|} | |||
====Make c3.2 working stocks==== | |||
Final concentration of each oligo in working stocks is 250 nM. | |||
{| {{table}} | |||
| align="center" style="background:#f0f0f0;"|'''working stock''' | |||
| align="center" style="background:#f0f0f0;"|'''description''' | |||
| align="center" style="background:#f0f0f0;"|'''pre-working stocks''' (according to [[IGEM:Harvard/2006/Container_Design_3#Working_stocks|chart]]) | |||
| align="center" style="background:#f0f0f0;"|'''water''' | |||
| align="center" style="background:#f0f0f0;"|'''total''' | |||
|- | |||
|c3.2.Db||-latches<br>+aptamers_in||1 (94 {{ul}}), 2 (28 {{ul}}), 3 (30 {{ul}}), 5 (3 {{ul}}), 6b (3 {{ul}}), 8 (2 {{ul}}), 9 (0 {{ul}})||40 {{ul}}||200 {{ul}} | |||
|- | |||
|c3.2.Eb||-latches<br>+aptamers_out||1 (94 {{ul}}), 2 (28 {{ul}}), 3 (30 {{ul}}), 4 (3 {{ul}}), 7b (3 {{ul}}), 8 (2 {{ul}}), 9 (0 {{ul}})||40 {{ul}}||200 {{ul}} | |||
|- | |||
|c3.2.H.core||+latch2<br>+aptamers_in||1 (94 {{ul}}), 2 (28 {{ul}}), 3 (30 {{ul}}), 5 (3 {{ul}}), 6b (3 {{ul}}), 10 (2 {{ul}}), 12 (2 {{ul}}), 13 (0 {{ul}}), 15 (2 {{ul}})||36 {{ul}}||200 {{ul}} | |||
|- | |||
|c3.2.H.latches||+latch2<br>+aptamers_in||16 (4 {{ul}})||-||- | |||
|- | |||
|c3.2.I.core||+latch2<br>+aptamers_out||1 (94 {{ul}}), 2 (28 {{ul}}), 3 (30 {{ul}}), 4 (3 {{ul}}), 7b (3 {{ul}}), 10 (2 {{ul}}), 12 (2 {{ul}}), 13 (0 {{ul}}), 15 (2 {{ul}})||36 {{ul}}||200 {{ul}} | |||
|- | |||
|c3.2.I.latches||+latch2<br>+aptamers_out||16 (4 {{ul}})||-||- | |||
|- | |||
|c3.2.Lb||-latches<br>+aptamers_all||1 (94 {{ul}}), 2 (28 {{ul}}), 3 (30 {{ul}}), 6b (3 {{ul}}), 7b (3 {{ul}}), 8 (2 {{ul}}), 9 (0 {{ul}})||40 {{ul}}||200 {{ul}} | |||
|} | |} | ||
Revision as of 14:44, 24 July 2006
Precipitation tests
- Made 10 mL solution with Dr. Shih's protocol.
- Tested (which DNA?) with the PEG precipitation protocol.
Streptavidin binding assay
- Double asterisks indicate need for purified nanostructures
- Total volume for each reaction: 13 μL
| lane | streptavidin (0.5 μM) (μL) | p7308 scaffold (44 nM) (μL) | oligos (250 nM) (μL) | nanostructures (10 nM) (μL) | water (μL) | 10x Tris-gly loading dye (μL) |
| 2 | 1.0 | - | - | - | 12.0 | 2 |
| 3 | - | 11.4 | - | - | 1.6 | 2 |
| 4 | 1.0 | 11.4 | - | - | 0.6 | 2 |
| 5 | - | - | 4.0 (a b-working stock) | - | 9.0 | 2 |
| 6 | 1.0 | - | 4.0 (same b-working stock) | - | 8.0 | 2 |
| 7 | 1.0 | - | - | 10.0** (3.2.A) | 2.0 | 2 |
| 8 | 1.0 | - | - | 10.0** (3.2.Eb or 4.0 Ib | 2.0 | 2 |
| 9 | 1.0 | - | - | 10.0** (3.2.Ib or 4.0.Ib) | 2.0 | 2 |
| 10 | 1.0 | - | - | 10.0** (3.2.Db or 4.0.Db) | 2.0 | 2 |
| 11 | 1.0 | - | - | 10.0** (3.2.Hb or 4.0.Hb) | 2.0 | 2 |
| 12 | 1.0 | - | - | 10.0** (3.2.Lb) | 2.0 | 2 |
Make c3.2 working stocks
Final concentration of each oligo in working stocks is 250 nM.
| working stock | description | pre-working stocks (according to chart) | water | total |
| c3.2.Db | -latches +aptamers_in |
1 (94 μL), 2 (28 μL), 3 (30 μL), 5 (3 μL), 6b (3 μL), 8 (2 μL), 9 (0 μL) | 40 μL | 200 μL |
| c3.2.Eb | -latches +aptamers_out |
1 (94 μL), 2 (28 μL), 3 (30 μL), 4 (3 μL), 7b (3 μL), 8 (2 μL), 9 (0 μL) | 40 μL | 200 μL |
| c3.2.H.core | +latch2 +aptamers_in |
1 (94 μL), 2 (28 μL), 3 (30 μL), 5 (3 μL), 6b (3 μL), 10 (2 μL), 12 (2 μL), 13 (0 μL), 15 (2 μL) | 36 μL | 200 μL |
| c3.2.H.latches | +latch2 +aptamers_in |
16 (4 μL) | - | - |
| c3.2.I.core | +latch2 +aptamers_out |
1 (94 μL), 2 (28 μL), 3 (30 μL), 4 (3 μL), 7b (3 μL), 10 (2 μL), 12 (2 μL), 13 (0 μL), 15 (2 μL) | 36 μL | 200 μL |
| c3.2.I.latches | +latch2 +aptamers_out |
16 (4 μL) | - | - |
| c3.2.Lb | -latches +aptamers_all |
1 (94 μL), 2 (28 μL), 3 (30 μL), 6b (3 μL), 7b (3 μL), 8 (2 μL), 9 (0 μL) | 40 μL | 200 μL |
