Subject M5 brain lesion section 2
Immunoglobulin (Ig) variable region heavy chain libraries were constructed from an autopsy sample of a human subject with clinically defined multiple sclerosis. Separate libraries were generated for three non-consecutive sections of a white matter lesion. RNA was reverse transcribed into cDNA using gene-specific primers mapping to the constant region of the Ig heavy chain. Resultant cDNA was tagged with 15 nucleotide unique molecular identifiers (UIDs) and amplified by PCR in a multiplex reaction using primer sets for all IGHV (n=45) and constant regions (n=6). A sequence processing pipeline using pRESTO (Vander Heiden and Yaari et al, 2014. Bioinformatics 30(13):1930-2) is available online at http://clip.med.yale.edu/papers/Stern2014STM.
Yale
MS23