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Advice on getting good PacBio libraries

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  • Advice on getting good PacBio libraries

    Hi everyone,

    We are having issues getting good quality PacBio libraries from Bacteria and microalgae. Specifically, we got low yield (76 MB) and low length (5,800 bp) from several libraries run on separate occasions. Other samples run in parallel had good stats, so the problem was not in the instrument and likely not in the library prep at our supplier.

    The DNA samples looked OK on a gel before and after shearing, but the yields were lower than expected after size selection from library prep.

    Has anyone experienced difficulty in getting the appropriate DNA for PacBio to work well? We could only get one out of four libraries to sequence well... Any advice would be useful.

    One point to consider is that we sent the samples using GenTegra tubes rather than on dry ice. Does anyone have experience with these tubes for PacBio?