Ex Agarose Gels; Non-Denaturing Conditions; Denaturing Agents; Denaturing Conditions - Thermo Scientific Invitrogen E-Gel G8100 User Manual

Running RNA Samples on E-Gel
™
E-Gel EX Agarose Gels can be used to run RNA samples. RNA can be run under denaturing or non-denaturing
conditions. Use non-denaturing conditions only when checking for RNA quality, where accurately determining size
is not critical.
Important: Using other denaturing agents like Glyoxal, Formaldehyde, or Urea results in very poor separation and
band morphology on E-Gel
It is not recommended to run samples that were loaded with RNA loading buffer on the same gel as samples that are
loaded with water.
•
Non- Mix RNA sample with RNase-free water such that the final volume is 20 μL.
•
Do not heat. Load the entire sample onto the E-Gel
denaturing
•
Run RNA using the E-Gel
conditions
Denaturing
The only denaturing agent that is compatible with the E-Gel
Lower concentrations are also acceptable.
agents
Denaturing
There are two methods for denaturing your RNA sample to run on an E-Gel
conditions
Method 1
1. Mix RNA (250 ng–2 μg) sample with formamide (to 50–95%) such that the final volume is 20 μL.
2. Heat samples at 65°C for 5 minutes to denature RNA.
3. Place samples on ice immediately after heating.
4. Load entire sample onto E-Gel
5. Run RNA using the E-Gel
Method 2
1. Mix RNA (250 ng–2 μg) sample with RNAse-free water or loading buffer such that the final
volume is 20 μL.
2. Heat samples at 65°C for 5 minutes to denature RNA
46
™ EX.
™
™

EX Agarose Gels

™
™
EX 1−2% program for 10 minutes.
™ EX.
EX 1−2% program for 10 minutes.
E-Gel Power Snap Electrophoresis System User Guide
™
Appendix E
EX.
™ EX system is Formamide, 50–95%.
™ EX Agarose Gel.