i have sorted anti-NP specific plasma cells from bone marrow of C57BL/6 mice at certain times after immunization with variable counts and isolated total RNA using TRIZOL method for RT-PCR using gene specific primer for C gamma 1 constant region of antibody heavy chain gene V186.2 and after that i used 2 rounds of nested PCR to amplify cDNA but i didn't get bands in gel in most of samples.
my main reference is : Affinity Maturation Develops in Two Stages of Clonal Selection 1998
Sabine Strehl
May I ask you how many cells you sorted and did you sort the cells directly into Trizol? And did you measure the RNA yield?
A typical mammalian cell contains 10–30 pg total RNA, so you can calculate the theoretical yield of RNA from a certain number of cells.
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