Daniel:Notebook/ComboLock/Polyacrylamide: Difference between revisions
>Djacobse (Created page with "=Polyacrylamide Gel Recipes for Combo Lock= Back to Notebook PA gel creation recipes. ==APS and TEMED== Based on the [[http://arep.med.harvar...") |
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Based on the [[http://arep.med.harvard.edu/polony/polony_protocols/bind_silane.htm protocol] from George Church's lab for PA Gel creation on glass slides. | Based on the [[http://arep.med.harvard.edu/polony/polony_protocols/bind_silane.htm protocol] from George Church's lab for PA Gel creation on glass slides. | ||
This recipe is done on the Drop Seq device and involves using APS and TEMED, which is a fast reaction done at room temperature. | This recipe is done on the Drop Seq device and involves using APS and TEMED, which is a fast reaction done at room temperature. As such, you make two different solutions, one with the acrylamide and bis monomers, the other with APS and TEMED. These are combined at the first cross-junction in the drop seq device. | ||
'''Acrylamide/Bis Solution''' | |||
This solution should also include sample, when using a sample. Fill the rest (after acylamide, bis, and sample) with water. | |||
{| class="wikitable" <hiddentext>generated with [[:de:Wikipedia:Helferlein/VBA-Macro for EXCEL tableconversion]] V1.8</hiddentext> | {| class="wikitable" <hiddentext>generated with [[:de:Wikipedia:Helferlein/VBA-Macro for EXCEL tableconversion]] V1.8</hiddentext> | ||
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| align="center" | | | align="center" | | ||
| align="center" | 500 | | align="center" | 500 | ||
|- style="background-color:#D9D9D9;font-size:12pt" valign="bottom" | |||
|style="font-weight:bold" height="15" | Total | |||
| align="center" | | |||
| align="center" | | |||
|style="font-weight:bold" align="center" | 1000 | |||
|} | |||
'''APS/TEMED''' Solution | |||
Mixed at the first cross junction in the Drop Seq device (must be added before oil/droplet formation). APS/TEMED is the initializer/accelerator, and therefore must be combined as late as possible before droplet formation to prevent clogging. | |||
{| class="wikitable" <hiddentext>generated with [[:de:Wikipedia:Helferlein/VBA-Macro for EXCEL tableconversion]] V1.8</hiddentext> | |||
|- style="background-color:#92CDDC;font-size:12pt;font-weight:bold" align="center" | |||
| width="116" height="41" | Reagent | |||
| width="90" | Stock Conc | |||
| width="137" | Final Conc | |||
| width="85" | Volume (uL) | |||
|- style="font-size:12pt" | |||
| height="15" valign="bottom" | Ammonium Persulfate | |||
| align="center" valign="bottom" | 5% | |||
| align="center" valign="bottom" | 0.2% (0.1%) | |||
| align="center" valign="bottom" | 40 | |||
|- style="background-color:#D9D9D9;font-size:12pt" | |||
| height="15" valign="bottom" | TEMED | |||
| align="center" valign="bottom" | 100% | |||
| align="center" valign="bottom" | 0.2% (0.1%) | |||
| align="center" valign="bottom" | 2 | |||
|- style="font-size:12pt" valign="bottom" | |||
| height="15" | dH20 | |||
| align="center" | | |||
| align="center" | | |||
| align="center" | 958 | |||
|- style="background-color:#D9D9D9;font-size:12pt" valign="bottom" | |- style="background-color:#D9D9D9;font-size:12pt" valign="bottom" |
Revision as of 17:27, 12 March 2016
Polyacrylamide Gel Recipes for Combo Lock
PA gel creation recipes.
APS and TEMED
Based on the [protocol from George Church's lab for PA Gel creation on glass slides.
This recipe is done on the Drop Seq device and involves using APS and TEMED, which is a fast reaction done at room temperature. As such, you make two different solutions, one with the acrylamide and bis monomers, the other with APS and TEMED. These are combined at the first cross-junction in the drop seq device.
Acrylamide/Bis Solution
This solution should also include sample, when using a sample. Fill the rest (after acylamide, bis, and sample) with water.
Reagent | Stock Conc | Final Conc | Volume (uL) |
Acrylamide | 40% | 18% (9%) | 450 |
Acrylamide/Bis | 40% (19:1) | 1.9%:0.1% (0.95%:0.05%) | 50 |
dH20 | 500 | ||
Total | 1000 |
APS/TEMED Solution
Mixed at the first cross junction in the Drop Seq device (must be added before oil/droplet formation). APS/TEMED is the initializer/accelerator, and therefore must be combined as late as possible before droplet formation to prevent clogging.
Reagent | Stock Conc | Final Conc | Volume (uL) |
Ammonium Persulfate | 5% | 0.2% (0.1%) | 40 |
TEMED | 100% | 0.2% (0.1%) | 2 |
dH20 | 958 | ||
Total | 1000 |