DNA Purifi cation Protocol for 4 ml Saliva Samples Cell Lysis 1. Incubate Oragene•DNA/Saliva samples at 50ºC in a water incubator for a minimum of 1 hour or in an air incubator for a minimum of 2 hours. 2. Transfer 4 ml lysate sample (2 ml saliva plus 2 ml Oragene•DNA-preserving solution) to a 15 or 50 ml centrifuge tube. 3. Add 1 ml Cell Lysis Solution and 25 μl Gentra RNase A Solution (4 mg/ml). Vortex on high speed for 10 seconds to mix sample and incubate 10 minutes at room

Ready; Catalysis Intro Helpful Texts: Principles and Applications of Organotransition Metal Chemistry, Collman, Hegedus, Norton, Fink The Organometallic Chemistry of the Transition Metals, Crabtree; Prof. M-Christina White’s lecture notes: http://www.courses.fas.harvard.edu/~chem153/ (many of my notes are taken from these, with permission) Advanced Synthesis and Catalysis, Spring 2011 Or: How to make more molecules more complex more quickly with less effort/material/waste

1 Napierala Lab protocol for culturing human iPSCs (May 2022) For details, please refer to “Maintenance of Human Pluripotent Stem Cells in mTeSR™1” Technical Manual (STEMCELL Technologies). Necessary Reagents Reagent Company Catalog Number BD MATRIGEL HESC-QUALIFIED Fisher Scientific 08-774-552 mTeSRTM1 Complete Kit STEMCELL Technologies 85850 Cryostor CS10 STEMCELL Technologies 07930 Dispase 1U/mL, 100mL STEMCELL Technologies 07923 Stemolecule Y27632 Stemgent 04-0012

Reactivity Umpolung-1 Ready Reactivity Umpolung: reversal of normal polarity electrophiles become nucleophiles nucleophiles become electrophiles Outline: 1. electrophilic heteroatoms 2. acyl anions 3. homoenolates 4. the cyclopropane trick 5. sulfone chemistry Normal reactivity: X x = heteroatom d = donor (Nu-) a = acceptor (E+) d a d a d X X X X or 1 3 1 5 How to make: X X or X X 1 2 1 4 complimentary

1 Napierala Lab protocol for human skin fibroblast culture (May 2022) Necessary Reagents Reagent Company Catalog Number DMEM, high glucose Gibco/Life Technologies 11965-092 Fetal bovine serum Hyclone SH30910.03 MEM Non-essential amino acids (NEAA) Gibco/Life Technologies 11140-050 0.25% Trypsin-EDTA, phenol red Gibco/Life Technologies 25200-056 DPBS (1X) Gibco/Life Technologies 14190-144 Fibroblast Medium recipe: DMEM, high glucose 15% fetal bovine serum (FBS

Advanced Synthesis and Catalysis, 2006 Unless noted, class is 9:00-10:30, mon and wed, L4.14 Date Topic Lecturer 8-22 Intro to catalysis Ready 8-24 Organometallics: structure/reactivity-1 Ready 8-29 Organometallics: structure/reactivity-2 Ready 8-31 Kinetics and mech-1 Ready 9-5 Kinetics and mech-2 Ready 9-7 Kinetic resolutions Ready 9-12 Nonlinear effects Ready 9-14 Addns to pi bonds-1 Ready 9-19 Addns to pi bonds-2 Ready 9-21 Addns to pi bonds-3 Ready 9-26 Conjugate addition-1 Ready 9-28

1 Sendai virus reprogrammed iPSC Lines Line Clone# Sex GAA repeat number in fibroblasts GAA repeat number in iPSCs Earliest passage Karyotype data Karyotype passage 1 CTRL 3348 6 M Control size Control size P5 Yes, normal P11 2 CTRL 7522 5 F Control size Control size P9 Yes, normal P15 3 CTRL 6718 1 M Control size Control size P2 Yes, normal P4 4 CTRL 6718 6 M Control size Control size P2 Yes, normal P4 5 CTRL 6719 1 F Control size Control size

1 FRDA fibroblasts Short alleles <499 repeats Medium alleles 500 – 999 repeats Long alleles >1000 repeats B - indicates line established from a second, independent skin biopsy of the same individual Cell Line Sex Sampling Age Age of Onset GAA repeats Allele 1 Allele 2 4 M 30 11 597 848 21 M 35 8 622 738 50 M 33 13 353 616 66 M 70 41 110 590 68 F 21 7 570 1200 86 F 30 18 367 780 88 F 50 16 520 520 90 F 29 7 556 556 90B F 33 7 570 570 130 M 56 41

L E C T U R E S E R I E S CARDIOMETABOLIC Dr. Vaduganathan received his medical degree from Northwestern University, where he also obtained a master’s degree of public health. He completed residency training at Massachusetts General Hospital and fellowship training in cardiovascular medicine at Brigham and Women’s Hospital. His research is centered on drug development, clinical trials, and the implementation of cardio-renal-metabolic therapies. Dr. Vaduganathan has authored or co-authored