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==APS and TEMED== 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. 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> |- style="background-color:#92CDDC;font-size:12pt;font-weight:bold" align="center" | width="116" height="49" | Reagent | width="90" | Stock Conc | width="137" | Final Conc | width="85" | Volume (uL) |- style="font-size:12pt" | height="15" valign="bottom" | Acrylamide | align="center" align="center" valign="bottom" | 40% | align="center" | 18% (9%) | align="center" valign="bottom" | 450 |- style="background-color:#D9D9D9;font-size:12pt" | height="15" valign="bottom" | Acrylamide/Bis | align="center" valign="bottom" | 40% (19:1) | align="center" | 1.9%:0.1% (0.95%:0.05%) | align="center" valign="bottom" | 50 |- style="font-size:12pt" valign="bottom" | height="15" | dH20 | align="center" | | align="center" | | 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="font-weight:bold" height="15" | Total | align="center" | | align="center" | |style="font-weight:bold" align="center" | 1000 |}
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