Hello all,
I am preparing a library of a selected number of specimens (20 ind) following the ddRad seq protocol (Peterson et al. 2013). I am having difficulties because I start with 500ng of Dna but I loose so much DNA that I end to get very low concentration before the final PCR.
In particular I loose more than 50% of DNA every beads wash or pooling. I using a concentration of 3X beads. Do you know if I am supposed to loose so much?
How many max PCR cycles may I run whitout increasing the PCR errors?
thank you
I am preparing a library of a selected number of specimens (20 ind) following the ddRad seq protocol (Peterson et al. 2013). I am having difficulties because I start with 500ng of Dna but I loose so much DNA that I end to get very low concentration before the final PCR.
In particular I loose more than 50% of DNA every beads wash or pooling. I using a concentration of 3X beads. Do you know if I am supposed to loose so much?
How many max PCR cycles may I run whitout increasing the PCR errors?
thank you
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