Poly l lysine pre coated clear bottom black walled 96 well plates

Telomere length measurement in peripheral blood mononuclear cells was performed as previously described [19 (link)]. Briefly, blood samples were thawed quickly and re-suspended in complete RPMI media and plated in poly-L-lysine pre-coated clear bottom black-walled 96-well plates (Greiner, Kremsmünster, Upper Austria, Austria). Samples were analyzed in duplicates. To convert telomeres fluorescence values into kb, we used standard cell lines with stable telomere length: L5178Y-R (79.7 kb), HeLa1211 (21.11 kb), Jurkat (11.5 kb), S (10.3 kb), K562 (6.5 kb), and HeLa R (6.03 kb). Images were acquired on an Opera High Content Screening System (PerkinElmer, Inc., Waltham, Massachusetts, USA) and analyzed with Acapella Image analysis software (PerkinElmer, Inc.).

To perform telomere length measurement in mouse peripheral blood mononuclear cells, blood samples were thawed quickly and re-suspended in complete RPMI media and plated in poly-L-lysine pre-coated clear bottom black-walled 96-well plates (Greiner, Kremsmünster, Upper Austria, Austria). Samples were analyzed in duplicates. The High-throughput Q-FISH (HT Q-FISH) protocol was performed as described by Canela et al. [62 (link)]. To convert telomeres fluorescence values into kb, we used standard cell lines with stable telomere length: L5178Y-R (79.7 kb), HeLa1211 (23.8 kb), Jurkat (11.5 kb), S (10.3 kb), K562 (6.5 kb), and HeLa R (6.03 kb) Images were acquired on an Opera High Content Screening System (PerkinElmer, Inc., Waltham, Massachusetts, USA) and analyzed with Acapella Image analysis software (PerkinElmer, Inc.).

To perform a longitudinal study on telomere length dynamics in mouse peripheral white blood cells, blood samples were collected as described in the section above at the indicated time points after AAV9 injection. Blood samples were processed with erythrocyte lysis using Buffer EL (QIAGEN) and frozen in 10% DMSO/FBS. Prior to proceeding with the protocol, blood samples were defrozen and plated in poly‐L‐lysine precoated clear bottom black‐walled 96‐well plates (Greiner, Kremsmünster, Upper Austria, Austria). Samples were analyzed in duplicate. The HT Q‐FISH protocol was performed as described in Canela et al. (2007). To convert telomeres fluorescence values into kb, we used standard cell lines with stable telomere length: L5178Y‐R (79.7 kb), HeLa1211 (23.8 kb), and CCRF‐CEM (7.5 kb). Images were acquired on an Opera High Content Screening System (PerkinElmer, Inc., Waltham, Massachusetts, USA) and analyzed with Acapella Image analysis software (PerkinElmer, Inc.).