Running Rna Samples On E-Gel ™ Ex Agarose Gels; Denaturing Agents; Prepare Ladder; Denature Sample - Thermo Scientific invitrogen E-Gel G8121ST User Manual

Running RNA Samples on E-Gel
E-Gel EX Agarose Gels can be used to run RNA samples. RNA is separated on the gel under denaturing conditions.
™
E-Gel EX Gel Type
™
1%
2%
Denaturing The only denaturing agent that is compatible with the E-Gel
formamide concentration in a prepared sample should be 50%.
agents
Important: Using other denaturing agents like glyoxal, formaldehyde, or urea results in very poor
separation and band morphology on E-Gel
Prepare Mix 500 ng of RNA ladder with formamide (50% final concentration) in a final volume of 20 μL.
ladder
Denature 1. Mix RNA (75 ng to 1.5 μg) sample with formamide (to 50% final concentration) in a final volume
of 20 μL.
sample
2. Load the entire sample (20 μL) into one well of an E-Gel
3. Fill any remaining empty wells with 20 μL of water or E-Gel
10482055).
4. Run the RNA using the E-Gel
Typical Universal Human Reference RNA (UHRR) total RNA was denatured with formamide without
heating and run on E-Gel
result of
RNA
Images were captured using the E-Gel
separation
to cool down for 5 minutes after being run for better sensitivity.
on E-Gel
™
EX agarose
gels
Lane M: Millennium
Lanes 1−9: UHHR; 75 ng, 100 ng, 150 ng,
E-Gel Power Snap Electrophoresis System User Guide
™
Recommended RNA ladder
Millennium
™
Century -Plus RNA Ladder (Cat. No. AM7145)
™
EX 1−2% program for 10 minutes.
™
EX agarose gels.
™
E-Gel EX 1% agarose gel
™
RNA Marker, 500 ng
™
200 ng, 250 ng, 500 ng, 750 ng,
1000 ng, 1500 ng
EX Agarose Gels
™
RNA Marker (Cat. No. AM7150)
EX system is formamide. The final
™
EX agarose gels.
™
EX agarose gel.
™
Power Snap Camera (Cat. No. G8200). Gels were allowed
™
E-Gel
Lane M: Century
Lanes 1−9: UHHR; 75 ng, 100 ng, 150 ng,
200 ng, 250 ng, 500 ng, 750 ng,
1000 ng, 1500 ng
Appendix E
E-Gel
™
18S–28S
5S–18S
Sample Loading Buffer (Cat. No.
™
EX 2% agarose gel
™
Plus RNA Ladder, 500 ng
™-
NGS
47