Dear Scientist,
I am looking for constructive feedback on the possible causes of pale shady band below the desired band on agarose gel. Kindly refer to the photo attached and you may find pale band below the desired band (353bp).
The gel was 1.25% and ran for 50 minutes. I don't think the pale shady band was caused by contamination because my non-template control (NTC) showed no any band.
Looking forward for any suggestion to overcome this. I am planning to send my PCR products for sanger sequencing and will the shady band affect the sequencing outcome?
Thank you.
Dear Zx, maybe there is an insertion sequence or mutations on your amplified product. Sanger sequencing of your PCR products and positive control could elucidate this problem.
Best regards.
Hi Mert Sudagidan , thanks for the comment. If there is an insertion sequence, the amplified product should have bigger size (>383bp) right? The pale band below the amplified product seems to have smaller size.
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