ena-STUDY-Centro de investigaciones oncologicas, Fundacion Cancer-16-11-2020-19:06:21:869-462
miRNome profiling of LSC-enriched CD34+CD38-CD26+ fraction in Ph+ CML-CP from Argentinean patients
Small-RNA sequencing of stem cell-enriched fractions from Chronic phase - Chronic myeloid leukemia patients at diagnosis, before treatment. Fractions were obtained by FACS from CD34+CD38-CD26+ (BCR-ABL1+) vs. CD34+CD38-CD26- (BCR-ABL1-) populations from CML-CP patients. Small-RNA sequecing was performed also in samples from healthy donors: CD34+CD38-/dim (normal hematopoietic stem cells) and CD34+CD38+ (progenitors). RNAs extracted from different samples were combined in order to increase RNA yield before NGS-library preparation. The entire content was used for library preparation (<140 ng for LSC, CML HSC and HD HSC fractions, and 210 ng for HD progenitor fraction).Libraries for small-RNA sequencing were prepared using Truseq Small RNA kit (Illumina), following manufacturer's instructions (15 PCR cycles). Single-end sequencing was performed on HiSeq 2500 (Illumina).
Small-RNA sequencing of stem cell-enriched fractions from Chronic phase - Chronic myeloid leukemia patients at diagnosis, before treatment. Fractions were obtained by FACS from CD34+CD38-CD26+ (BCR-ABL1+) vs. CD34+CD38-CD26- (BCR-ABL1-) populations from CML-CP patients. Small-RNA sequecing was performed also in samples from healthy donors: CD34+CD38-/dim (normal hematopoietic stem cells) and CD34+CD38+ (progenitors). RNAs extracted from different samples were combined in order to increase RNA yield before NGS-library preparation. The entire content was used for library preparation (<140 ng for LSC, CML HSC and HD HSC fractions, and 210 ng for HD progenitor fraction).Libraries for small-RNA sequencing were prepared using Truseq Small RNA kit (Illumina), following manufacturer's instructions (15 PCR cycles). Single-end sequencing was performed on HiSeq 2500 (Illumina).