Team:Arizona State/Lab/Protocols/Gel visualization

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Latest revision as of 08:40, 27 September 2011

Protocols: Gel visualization

50 mL Gel Electrophoresis

Mix 1x TAE Buffer with 0.50g agarose in 250 mL Erlenmeyer flask. Swirl well.
Microwave solution until it begins to boil. Gently swirl the flask and repeat the microwaving process until the agarose is completely dissolved and the solution is clear.
Add 5 μL dye (SYBR green or ethidium bromide) and gently swirl the mixture. Let the solution sit for 5 minutes.
Carefully pour the solution into the 50 mL gel tray and add in the desired comb. Let sit for 30-45 minutes until gel solidifies.
Place gel tray in the -4C refrigerator for 10-15 minutes.
Remove gel tray from refrigerator, remove rubber ends, and place the gel in electrophoresis chamber with the comb towards the negative end.
Add in enough 1x TAE Buffer in the chamber to just barely create a thin film of buffer over the gel.
Add 2 μL of each DNA sample to a strip of parafilm. Add 8 μL of loading buffer to each of those 2 μL samples.
Add the necessary DNA ladder to the first well in the gel.
Add each 10 μL mix of buffer and DNA into individual wells in the gel.
Attach the electrophoresis chamber lid onto the chamber and plug into the power source.
Set the power source to 100-110 V for fast visualization (or 80V for optimal separation and resolution) and let sit for approximately 30-45 minutes or until the dye marks are near the end of the gel.
Turn off the power source, remove the gel, and use ultraviolet light to visualize the DNA bands.

@@ Line 1: / Line 1: @@
-.	Mix 1x TAE Buffer with 0.50g agarose in 250 mL Erlenmeyer flask. Swirl well.
+{{:Team:Arizona State/Templates/main|title=Protocols: Gel visualization|content=
-.	Microwave solution until it begins to boil. Gently swirl the flask and repeat the microwaving process until the agarose is completely dissolved and the solution is clear.
+&nbsp;
-.	Add 5 μL SYBR green and gently swirl the mixture. Let the solution sit for 5 minutes.
+'''50 mL Gel Electrophoresis'''
-.	Carefully pour the solution into the 50 mL gel tray and add in the desired comb. Let sit for 30-45 minutes until gel solidifies.
-.	Place gel tray in the -4 o C refrigerator for 10-15 minutes.
+# Mix 1x TAE Buffer with 0.50g agarose in 250 mL Erlenmeyer flask. Swirl well.
-.	Remove gel tray from refrigerator, remove rubber ends, and place the gel in electrophoresis chamber with the comb towards the negative end.
+# Microwave solution until it begins to boil. Gently swirl the flask and repeat the microwaving process until the agarose is completely dissolved and the solution is clear.
-.	Add in enough 1x TAE Buffer in the chamber to just barely create a thin film of buffer over the gel.
+# Add 5 μL dye (SYBR green or ethidium bromide) and gently swirl the mixture. Let the solution sit for 5 minutes.
-.	Add 2 μL of each DNA sample to a strip of parafilm. Add 8 μL of loading buffer to each of those 2 μL samples.
+# Carefully pour the solution into the 50 mL gel tray and add in the desired comb. Let sit for 30-45 minutes until gel solidifies.
-.	Add the necessary Hyperladder to the first well in the gel.
+# Place gel tray in the -4C refrigerator for 10-15 minutes.
-.	Add each 10 μL mix of buffer and DNA into individual wells in the gel.
+# Remove gel tray from refrigerator, remove rubber ends, and place the gel in electrophoresis chamber with the comb towards the negative end.
-.	Attach the electrophoresis chamber lid onto the chamber and plug into the power source.
+# Add in enough 1x TAE Buffer in the chamber to just barely create a thin film of buffer over the gel.
-.	Set the power source to 100-110 V and let sit for approximately 30-45 minutes or until the dye marks are near the end of the gel.
+# Add 2 μL of each DNA sample to a strip of parafilm. Add 8 μL of loading buffer to each of those 2 μL samples.
-.	Turn of the power source, remove the gel, and use ultraviolet light to visualize the DNA bands.
+# Add the necessary DNA ladder to the first well in the gel.
+# Add each 10 μL mix of buffer and DNA into individual wells in the gel.
+# Attach the electrophoresis chamber lid onto the chamber and plug into the power source.
+#Set the power source to 100-110 V for fast visualization (or 80V for optimal separation and resolution) and let sit for approximately 30-45 minutes or until the dye marks are near the end of the gel.
+#Turn off the power source, remove the gel, and use ultraviolet light to visualize the DNA bands.
+}}