8561Tx WT
Donor-derived LSK cells regenerated in either WT or Wnt5a+/- recipients were sorted into lysis buffer composed of 0.2 % Triton X-100 (Sigma) and 2 U/µl of RNAse Inhibitor (Life Technologies). ERCC spike-in controls (Life Technologies) were added to the cell lysis mix at 1:5,000 dilution. cDNA was synthesized and pre-amplified from 3.5 µl of lysate as described elsewhere (Picelli et al., 2013). 500 pg of cDNA was used as input for tagmentation by the Nextera XT Sample Preparation Kit (Illumina), where a second amplification round was performed for 10 cycles. For each sample, 3 ng of final library was pooled
Ludwig-Maximilians University Munich
8561Tx WT