RNA was extracted from FACS sorted naïve, GC and post-GC memory B cells of human tonsils, as well as from human DLBCL cell lines using miRNeasy mini kit (Qiagen). cDNA was synthesized using miRCURY LNA RT kit (Qiagen) and quantified by SYBR green assay (miRCURY LNA SYBR Green PCR Kit, Qiagen). For detection of human mature miR-28a-5p, the following miRCURY LNA miR-28 primer (Qiagen; Cat# 2YP00204322) was used. U6 amplification (Qiagen; Cat#YP00203907) was used as normalization control. Amplification was performed in QuantStudio5-384 Real-Time PCR System (Applied Biosystems) equipment.
Do you have any questions about this protocol?
Post your question to gather feedback from the community. We will also invite the authors of this article to respond.