Supporting documents for PROseq samples. Please see the main PROseq page for more information.
Note that good permeablized cells are the single most important determinant of good PRO-seq libraries. Please read the permeabilization protocol carefully and execute it with care. For returning users, please note that the permeabilization protocol has recently been revised. The original and the revised protocols give cells of equal quality for nuclear run-on but we find the revised protocol is easier to execute.
PRO-seq samples should be submitted at a concentration of 1M cells / 100uL. If possible, please provide a single aliquot of 5M cells in 500uL. We will recount cells prior to library construction. If cells are ≤80% permeabilized it is unlikely that run-on reactions will perform well and we will be in touch about repreparing cells.