title: Illumina HiSeq 4000 sequencing; leChRO-seq for 2 pancreatic cancer patient-derived xenografts treated with Vehicle or Gemcitabine/Paclitaxel in mouse
design: leChRO-seq for 2 pancreatic cancer patient-derived xenografts treated with Vehicle or Gemcitabine/Paclitaxel in mouse
library_construction_protocol: Both PDXs are from adults. The tissue is Pancreas and it was the initial tumor site. PDX13 was established from surgically resected tumor tissue from a PDAC patient at the Ruhr-University Bochum Comprehensive Cancer Center. Informed and written consent was obtained. Approval by the ethics committee of the Ruhr University Bochum was obtained under Permission no. 3534-9, 3841-10, 16-5792. Experiments were performed according to the guidelines of the local Animal Use and Care Committees at the Ruhr University Bochum (8.87-50.10.32.09.018, 84-02.04.2012.A328  and 81-02.04.2017.A423). Establishment: Tumor tissues was implanted in NMRInu/nu mice (both flanks). Tumors were expanded and re-implanted for at least three generations. Treatment: 1-2mm of tumor tissue (early passage) were implanted subcutaneously in 5â€“10-week-old mice (NMRInu/nu). Treatment was started when tumors were approximately 100-200 mm3 in size. Mice were randomized into vehicle (09.% saline) or Gemcitabine/Paclitaxel  (100/30 mg/kg).cells were collected  and washed in PBS once, while 30mg of PDX were homogenized using Biomasher II (VWR). RNA was extracted using TRIzol 5 prime RBA adaptors were ligated using T4 RNA ligase. Phusion High Fidelity DNA Polymerase was used to PCR amplify the library, which was then submitted to a post-PCR cleanup using AMPure XP beads.