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Custom protocol sequencing gave 90% junk sequences. Rest is matching the human genome. Majority of junk sequences are repeats of adapter/primer, where the sequencing primer should bind. You can get a gist of the aligned sequences and the organization of the final library product in the attached pdf.
I am using RP1 and RPI1 primers with a PS (Phosphorothioate) bonds at the penultimate base. Both the primers are HPLC purified from a reputed supplier. Earlier I had a doubt that PS bonds are interfering cluster formation. However, techsupport clearly mentioned that this cannot be the case. Is it because of some contaminating primer from the other libraries, which were sequenced in the lane? Has anyone seen such sequences in their libraries?
Your input will be of great help! Thanks!
BB
Custom protocol sequencing gave 90% junk sequences. Rest is matching the human genome. Majority of junk sequences are repeats of adapter/primer, where the sequencing primer should bind. You can get a gist of the aligned sequences and the organization of the final library product in the attached pdf.
I am using RP1 and RPI1 primers with a PS (Phosphorothioate) bonds at the penultimate base. Both the primers are HPLC purified from a reputed supplier. Earlier I had a doubt that PS bonds are interfering cluster formation. However, techsupport clearly mentioned that this cannot be the case. Is it because of some contaminating primer from the other libraries, which were sequenced in the lane? Has anyone seen such sequences in their libraries?
Your input will be of great help! Thanks!
BB