Difference between revisions of "Team:Stony Brook/Notebook/Cancer-W7"
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<li>Didn't work</li> | <li>Didn't work</li> | ||
</ul> | </ul> | ||
| + | |||
| + | <h4> Nanodrop of Miniprepped Plamids: </h4> | ||
| + | <ul> | ||
| + | <table> | ||
| + | <tr> | ||
| + | <th> Tube # </th> | ||
| + | <th> Concentration (ng/ul) </th> | ||
| + | </tr> | ||
| + | <tr> | ||
| + | <td> 1 </td> | ||
| + | <td> 277.3 </td> | ||
| + | </tr> | ||
| + | <tr> | ||
| + | <td> 2 </td> | ||
| + | <td> # </td> | ||
| + | </tr> | ||
| + | <tr> | ||
| + | <td> 2 </td> | ||
| + | <td> # </td> | ||
| + | </tr> | ||
| + | <tr> | ||
| + | <td> 2 </td> | ||
| + | <td> # </td> | ||
| + | </tr> | ||
| + | </table> | ||
| + | </ul> | ||
| + | |||
<h5> PCR of Minprepped Plasmids </h5> | <h5> PCR of Minprepped Plasmids </h5> | ||
| Line 479: | Line 506: | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
| − | <td> </td> | + | <td> Q5 Polymerase </td> |
| − | <td> | + | <td> 0.25 uL </td> |
</tr> | </tr> | ||
<tr> | <tr> | ||
| − | <td> | + | <td> Q5 Polymerase </td> |
| − | <td> 0. | + | <td> 0.25 uL </td> |
</tr> | </tr> | ||
</table> | </table> | ||
</ul> | </ul> | ||
| − | <h4> PCR Settings | + | <h4> PCR Settings: </h4> |
| − | + | ||
<ul> | <ul> | ||
<table> | <table> | ||
| Line 501: | Line 527: | ||
<td> Initial Denaturation </td> | <td> Initial Denaturation </td> | ||
<td> 98 </td> | <td> 98 </td> | ||
| − | <td> | + | <td> 30s </td> |
</tr> | </tr> | ||
<tr> | <tr> | ||
<td> Denaturation </td> | <td> Denaturation </td> | ||
<td> 98 </td> | <td> 98 </td> | ||
| − | <td> | + | <td> 20s </td> |
</tr> | </tr> | ||
<tr> | <tr> | ||
<td> Annealing </td> | <td> Annealing </td> | ||
| − | <td> | + | <td> 56 </td> |
| − | <td> | + | <td> 15s </td> |
</tr> | </tr> | ||
<tr> | <tr> | ||
<td> Extension </td> | <td> Extension </td> | ||
<td> 72 </td> | <td> 72 </td> | ||
| − | <td> | + | <td> 30s </td> |
</tr> | </tr> | ||
<tr> | <tr> | ||
<td> Final Extension </td> | <td> Final Extension </td> | ||
<td> 72 </td> | <td> 72 </td> | ||
| − | <td> | + | <td> 60s </td> |
</tr> | </tr> | ||
</table> | </table> | ||
</ul> | </ul> | ||
| + | |||
| + | |||
Revision as of 18:33, 11 August 2016
I left you some of the commonly used templates down there under 8/8. Just fill them in as needed. I'll only really have wifi for a few hours on two days between Saturday and Thursday. If you need anything, message me on Facebook and I'll get back to you as soon as I can.
With the warmest love and regards,
~JOSHUA
Week 7: 8/8 - 8/14
PCR Table Template:
| Contents | ul | Phire | # |
|---|---|
| H2O | # |
| 50 uM F Primer | # |
| 50 uM R Primer | # |
| #ug/ul Construct | # |
PCR Settings Template:
| Phase | Temperature (°C) | Time (sec) |
|---|---|---|
| Initial Denaturation | 98 | # |
| Denaturation | 98 | # |
| Annealing | # | # |
| Extension | 72 | # |
| Final Extension | 72 | # |
Nanodrop Template:
| Content | Concentration (ng/ul) | 260/280 |
|---|---|---|
| Construct | # | # |
| Vector | # | # |
Ligation Template
| Content | ul |
|---|---|
| 10X T4 Buffer | # |
| Vector | # (# ng) | Construct | # (# ng) |
| H2O | # |
| Ligase | # |
Multiple Digests Template:
This
| Content | Experimental ul | Control ul |
|---|---|---|
| H2O | # | # |
| Cutsmart | # | # |
| EcoRI-HF | # | X |
| XbaI | # | X |
| DNA | # | # |
Is
| Content | Experimental ul | Control ul |
|---|---|---|
| H2O | # | # |
| Cutsmart | # | # |
| EcoRI-HF | # | X |
| XbaI | # | X |
| DNA | # | # |
A
| Content | Experimental ul | Control ul |
|---|---|---|
| H2O | # | # |
| Cutsmart | # | # |
| EcoRI-HF | # | X |
| XbaI | # | X |
| DNA | # | # |
Thing
| Content | Experimental ul | Control ul |
|---|---|---|
| H2O | # | # |
| Cutsmart | # | # |
| EcoRI-HF | # | X |
| XbaI | # | X |
| DNA | # | # |
8/8
Sequencing
| T1 | T2 |
|---|---|
| 5.34 uL H2O | 5.34 uL H2O |
| 1 uL CR-1 Primer 1 | 1 uL CR-1 Primer 2 | 1.66 uL DNA (301.6 ng/uL) | 1.66 uL DNA (301.6 ng/uL) |
Diluted BioBrick MiniPreps for PCR
| BioBrick Name | Concentration (ng/uL) | Contents (DNA + water) |
|---|---|---|
| K592009 | 76.9 | 1 uL + 6.69 uL water |
| B0015 | 51.7 | 1 uL + 4.17 uL water |
| K1033910 | 156.5 | 1 uL + 14.65 uL water |
| R0082 | 49.5 | 1 uL + 3.95 uL water |
| C0012 | 150.8 | 1 uL + 14.08 uL water |
| J61100 | 123.0 | 1 uL + 11.3 uL water |
| R0011 | 56.7 | 1 uL + 4.67 uL water |
PCR of each BioBrick parts (25 uL Q5 MasterMix rxns)
| Contents | uL |
|---|---|
| 10 uM VF2 primer | 1.25 uL |
| 10 uM VR primer | 1.25 uL |
| DNA | 0.5 uL |
| Q5 MM | 12.5 uL |
| Water | 19.5 uL |
8/9
Gel ran
Lanes:
- Ladder
- R0011
- B001
- K1033
- R008
- J61100
- C0012
- K592
- Didn't work
Nanodrop of Miniprepped Plamids:
| Tube # | Concentration (ng/ul) |
|---|---|
| 1 | 277.3 |
| 2 | # |
| 2 | # |
| 2 | # |
PCR of Minprepped Plasmids
| Contents | ul | Q5 Buffer | 5uL |
|---|---|
| H2O | 16.25 uL |
| 10 uM F Primer | 1.25 uL |
| 10 uM R Primer | 1.25 uL |
| Construct | 0.5 uL |
| Q5 Polymerase | 0.25 uL |
| Q5 Polymerase | 0.25 uL |
PCR Settings:
| Phase | Temperature (°C) | Time (sec) |
|---|---|---|
| Initial Denaturation | 98 | 30s |
| Denaturation | 98 | 20s |
| Annealing | 56 | 15s |
| Extension | 72 | 30s |
| Final Extension | 72 | 60s |
8/10
8/11
8/12
8/13
8/14
