Nucleic Acid Isolation
Blood Samples for RNA extractions
Total RNA will be extracted using the QIAsymphony PAXgene Blood RNA Kit (https://www.qiagen.com/us/shop/sample-technologies/rna/mirna/qiasymphony-paxgene-blood-rna-kit/#orderinginformation).
All blood samples (2.5ml) should be collected in PAXgene Blood RNA Tubes (https://www.preanalytix.com/products/blood/RNA/paxgene-blood-rna-tube). Immediately after blood collection, gently invert the PAXgene Blood RNA Tubes 8–10 times. Store the PAXgene Blood RNA Tubes upright at room temperature (18−25 ̊C) for a minimum of 2 hours and a maximum of 72 hours before transferring to freezer. The PAXgene Blood RNA Tubes can be stored at −20 ̊C and below. If tubes are to be kept at temperatures below −20 ̊C, freeze them first at −20 ̊C for 24 hours, then transfer them to −80 ̊C.
Tubes should be delivered to us frozen. Note that the frozen PAXgene Blood RNA Tubes are very fragile and break easily.
Tubes containing material for isolation must be clearly labeled with your DUGSIM order number and sample number (“4596_S001”). For human blood samples, please make sure to provide us with your IRB number to confirm that your project has the appropriate IRB approval.
Cell pellets for RNA extractions
Total RNA from cell pellets will be extracted using the Qiagen QIAsymphony RNA Kit (https://www.qiagen.com/us/shop/automated-solutions/qiasymphony-rna-kit/#orderinginformation).
Cell pellets should have between 1-3 million cells.
Cells should be pelleted in Sarstedt 2.0 mL tubes - cat # 72.693 or 72.608