Daniel:Notebook/ComboLock/Polyacrylamide

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Revision as of 17:28, 12 March 2016 by >Djacobse (→‎APS and TEMED)
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Polyacrylamide Gel Recipes for Combo Lock

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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

VA-044