![]() Following electrophoresis, visualize by staining in ethidium bromide solution (final concentration 0.5 μg/ml) or SYBR Green I. ![]() Follow the recommendations for loading described in the certificate of analysis of the ladders/markers (~ µg per 1 mm gel lane width) or in the Table.6 on p Insufficient or uneven staining. Troubleshooting Guide for Electrophoresis Insufficient amount of ladder was loaded. Low intensity of all or some of the bands Table.8. ELECTROPHORESIS Protocols and Recommendations for Electrophoresis electrophoresis problem 1 Low intensity of all or some bands 2 Smeared bands 3 Atypical banding pattern 4 Curved bands 5 remains in the gel well 6 Incorrect quantification data 1.1 Insufficient amount of loaded 2.1 degradation by nucleases 3.1 l marker not heated prior to loading 4.1 Gel incompletely immersed in buffer 5.1 Poorly-formed gel wells 6.1 Different loading conditions for ladder & sample 1.2 Insufficient or uneven staining 2.2 Improper electrophoresis conditions 3.2 Denatured 4.2 Low sample volume 5.2 Excess loaded 6.2 Incorrect ladder band chosen for quantification 1.3 run off of the gel 2.3 Gel shift effect 3.3 Different loading conditions for ladder & sample 4.3 Improper electrophoresis conditions 5.3 Contamination of the sample 6.3 Improper quantification method 1.4 diffusion in the gel 2.4 Excess loaded 3.4 Improper electrophoresis conditions 4.4 Bubbles or physical particles in wells or in the gel 5.4 Gel shift effect 6.4 Uneven or high background staining of the gel 1.5 masking by tracking dyes 2.5 High salt concentration in samples 3.5 Incorrect gel percentage or running buffer used 6.5 masking by tracking dyes 2.6 Poorly formed gel wells 3.6 Atypical migration due to sequence or structure 3.7 Gel shift effect 3.8 High salt concentration in samples Bulk quantities and custom formulations available upon request 447Ģ. 1 Troubleshooting Guide for Electrophoresis.
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