Illumina HiSeq 1500 sequencing; Bulk RNA-seq of archived Acute Myeloid Leukemia (AML) samples propagated in a mouse Xenograft model over several passages
Bulk RNA-seq of archived Acute Myeloid Leukemia (AML) samples propagated in a mouse Xenograft model over several passages
Cryopreserved AML-PDX cells have routinely been frozen as 10 Mio cells in 1mL of freezing medium (80% FBS, 20% DMSO) at -80 °C for archiving purposes. To avoid thawing these samples and thus harming or even destroying them, the frozen cell stocks were first transferred to dry ice under a cell culture hood. Next a sterile 1mm biopsy punch was used to punch the frozen cells in the vial and transfer the extracted cells to one well of a 96 well plate containing 100 µL RLTplus lysis buffer with 1% beta mercaptoethanol. To ensure complete lysis the lysate was mixed and snap frozen on dry ice. prime-seq RNA extraction using magnetic beads (see prime-seq protocol) The prime-seq protocol was performed as described here: https://www.protocols.io/view/prime-seq-s9veh66