Difference between revisions of "Team:NAIT Edmonton/Protocols"

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     <p><b><u>Mix</u></b></p>
 
     <p><b><u>Mix</u></b></p>
 
     <p> - 4.2ml DH20<br> - 2.5ml TRIS 4X Buffer.<br> - 3.3ml Polyacrylamide Solution.<br><br> 3) Mix and invert six times<br> 4) Add 33.3μl of APS (Ammonium Persulfate) into the falcon tube.<br> 5) Mix and invert six times<br> 6)Add 6.7μl of TEMED to the falcon tube.
 
     <p> - 4.2ml DH20<br> - 2.5ml TRIS 4X Buffer.<br> - 3.3ml Polyacrylamide Solution.<br><br> 3) Mix and invert six times<br> 4) Add 33.3μl of APS (Ammonium Persulfate) into the falcon tube.<br> 5) Mix and invert six times<br> 6)Add 6.7μl of TEMED to the falcon tube.
     <br> <b>NOTE:</b> Must be done in a fumehood. TEMED is toxic.<br> 7) Mix and invert six times. Solution will harden in roughly 15 minutes.<br> 8) Add solution in between glass plates using a pipette aid roughly 3/4 the way up the front plate.<br> 9) Fill remaining space with ethanol to ensure an even leveling of the gel.<br> 10) Once gel is solidified, remove the ethanol using a kim wipe ensuring the are in bewteen the plates is now dry.<br><br></p>
+
     <br> <b>NOTE:</b> Must be done in a fumehood. TEMED is toxic.<br><br> 7) Mix and invert six times. Solution will harden in roughly 15 minutes.<br> 8) Add solution in between glass plates using a pipette aid roughly 3/4 the way up the front plate.<br> 9) Fill remaining space with ethanol to ensure an even leveling of the gel.<br> 10) Once gel is solidified, remove the ethanol using a kim wipe ensuring the are in bewteen the plates is now dry.<br><br></p>
  
 
     <p style="font-size:16px"><b><u>Stacking Gel</u></b>:<br></p>
 
     <p style="font-size:16px"><b><u>Stacking Gel</u></b>:<br></p>
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     <p><b><u>Mix</u></b></p>
 
     <p><b><u>Mix</u></b></p>
 
     <p> - 3.05ml DH20<br> - 1.25ml Stacking Buffer Solution<br> - 650μl Polyacrylamide Solution.<br><br> 2)      Mix and invert six times.<br> 3) Add 25μl of APS ( Ammonium Persulfate ) into the falcon tube.<br> 4) Mix and invert six times.<br> 5) Add 5μl of TEMED to solution.  
 
     <p> - 3.05ml DH20<br> - 1.25ml Stacking Buffer Solution<br> - 650μl Polyacrylamide Solution.<br><br> 2)      Mix and invert six times.<br> 3) Add 25μl of APS ( Ammonium Persulfate ) into the falcon tube.<br> 4) Mix and invert six times.<br> 5) Add 5μl of TEMED to solution.  
     <br><b>NOTE:</b> Must be done in a fumehood. TEMED is toxic.<br> 6) Mix and invert six times. <br> 7) Fill in the remaining area in between the glass plates with the stacking solution and place comb inside.<br> 8) After gel is solidfied, remove from casting frame and place in plastic wrap. Store in the fridge. <br> READY FRO USE!<br><br></p>
+
     <br><b>NOTE:</b> Must be done in a fumehood. TEMED is toxic.<br><br> 6) Mix and invert six times. <br> 7) Fill in the remaining area in between the glass plates with the stacking solution and place comb inside.<br> 8) After gel is solidfied, remove from casting frame and place in plastic wrap. Store in the fridge. <br> READY FRO USE!<br><br></p>
  
 
   </div>
 
   </div>

Revision as of 22:18, 17 August 2015

Team NAIT 2015

Experimental Design

Go through our interactive experimental design flow chart! Many of our protocols are manufacturer specicfied; however, some are customized by us! PDFs of protocols can also be found

Theorizing our Sequences
Literature has shown certain proteins inherently stain in colour.
Looked up characteristics of said proteins
Isolated and identified the unique characteristics of said proteins so that we can manually write our own sequences and generate custom proteins.
Writing our Sequences
Went down to base pair level and wrote out our sequences with the defining characteristics identified in literature.
PCR
Digestion and Ligation
Transforming Bacteria
Validating the Transformation
Protein Isolation and Purification
SDS PAGE and Silver Staining