Table 3.
Reagents details.
| Antibodies used for immunocytochemistry/flow-citometry | |||
| Antibody | Dilution | Company Cat # and RRID | |
|
| |||
| Pluripotency Marker | Mouse anti-Oct-4, Alexa Fluor® 488 conjugate | 1:100 | Millipore Sigma, MAB4419A4, RRID:AB_2847875 |
| Pluripotency Marker | Mouse anti-Sox-2, Cy3 conjugate | 1:100 | Millipore Sigma, MAB4423C3, RRID:AB_2847876 |
| Pluripotency Marker | Mouse anti-Nanog, Alexa Fluor® 488 conjugate | 1:100 | Millipore Sigma, SCR078, MABD24A4, RRID:AB_2847877 |
| Mesoderm marker | smooth muscle actin | 1:200 | Invitrogen, A25531, RRID:AB_2651005 |
| Endoderm marker | α-fetoprotein | 1:200 | Invitrogen, A25530, RRID:AB_2651004 |
| Ectoderm marker | β-III tubulin | 1:200 | Invitrogen, A25532, RRID:AB_2651003 |
| Primers | |||
| Target | Forward/Reverse primer (5′-3′) | ||
|
| |||
| Mycoplasma testing | Mycoplasma | Forward primers: CGCCTGAGTAGTACGTTCGC; CGCCTGAGTAGTACGTACGC; TGCCTGAGTAGTACATTCGC; TGCCTGGGTAGTACATTCGC; CGCCTGAGTAGTATGCTCGC/ Reverse primers: GCGGTGTGTACAAGACCCGA; GCGGTGTGTACAAAACCCGA; GCGGTGTGTACAAACCCCGA |
|
| Mutation analysis for HT188 | PARK7 | TTCTGTGCTTTTGCCAGATG/TCTTTAGCAAGAGGGTGTGTTG | |
| CRISPR diagnostic primers | Donor oligo | TGGGCTCAAGCAATTTTTCTA/ATGGAGCGAGACTCCATCT | |
| gRNA for A18945 | PARK7 mutation | caccgCTTAGGTCCTACTGCTCTGT/ aaacACAGAGCAGTAGGACCTAAGc | |
| Donor oligo | TGCGATTTTTTAAACATGGGCTTTTCTATATCTGCACTTAGAT | ||
| CTTTTTATTTTTATTCTTAGGTCCTACTCTCTGTTGGCTCATGA | |||
| AATAGGTTTTGGAAGTAAAGTTACAACACACCCTCTTGCTAA | |||
| AGACAAAATGATGAATGGAGGTAAGTATATGCTTGTTTTTGT | |||
| TTGTTTGTTTGTTTTTTGAGATGGAGTC | |||