I get a mutation by CRISPR, but when I do the sequencing I always get the results with poly peaks. How can I know the type of my mutation? The poly peaks always present from my target, and shows 3 or 4 different bases.
For a good separation of the sequences you can pcr across the region and clone the pcr product then sequence clones. Otherwise look at the sequence and subtract the normal sequence and any strong peaks that remain are probably the ones that you introduced. If there were many polypeaks then possibly you have introduced an insertion/deletion of a base
You can use the webpage which called "TIDE" to alyze the type of your mutation or as Paul suggested the best way is to clone it (I used PGEM-T easy vector) do blue/white selection and send the independent clones (white colonies or light blue ones) for sequencing.
Just in case I put the web address of TIDE
https://tide.nki.nl
By the way when you send your clones for sequencing I suggest also to use T7 or SP6 universal primers and sequence both strand. You can also send with the forward and reverse primers that you used for your pcr :)
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