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After cells are isolated from tissue or differentiated from pluripotent precursors, the resulting population needs to be characterized to confirm whether the target population has been obtained. The table below lists PCR primers that can be used in end point polymerase chain reactions (PCR) to detect the expression of specific genes for characterizing neural stem cells (NSCs) and their sublineages. Suitability of the listed primers should be determined on an experimental basis, and optimization of reaction conditions, including redesign of the primer sequences themselves, is recommended to ensure proper lineage characterization.
| Target | Primer | Sequence | Tm (°C) | Amplicon size (bp) | Intron size (bp) |
|---|---|---|---|---|---|
| Neural stem cells | SOX1-F | GCGGAAAGCGTTTTCTTG | 53.0 | 406 | No Intron |
| SOX1-R | TAATCTGACTTCTCCTCCC | 50.2 | |||
| SOX2-F | ATGCACCGCTACGACGTGA | 59.3 | 437 | No Intron | |
| SOX2-R | CTTTTGCACCCCTCCCATTT | 56.0 | |||
| NESTIN-F | CAGCGTTGGAACAGAGGTTGG | 58.6 | 389 | 1,142 | |
| NESTIN-R | TGGCACAGGTGTCTCAAGGGTAG | 60.7 | |||
| Oligodendrocytes | MAG-F | TCTGGATTATGATTTCAGCC | 49.7 | 366 | 159 |
| MAG-R | GCTCTGAGAAGGTGTACTGG | 54.7 | |||
| OSP-F | ACTGCTGCTGACTGTTCTTC | 55.1 | 283 | 5,714 | |
| OSP-R | GTAGAAACGGTTTTCACCAA | 50.8 | |||
| Astrocytes | ALDH1L1-F | TCACAGAAGTCTAACCTGCC | 55.5 | 398 | 21,837 |
| ALDH1L1-R | AGTGACGGGTGATAGATGAT | 54.4 | |||
| GFAP-F | GTACCAGGACCTGCTCAAT | 55.0 | 321 | 2,989 | |
| GFAP-R | CAACTATCCTGCTTCTGCTC | 55.3 | |||
| Neurons | MAP2-F | CCACCTGAGATTAAGGATCA | 55.1 | 482 | 11,798 |
| MAP-R | GGCTTACTTTGCTTCTCTGA | 55.0 | |||
| ChAT-F | ACTGGGTGTCTGAGTACTGG | 55.0 | 451 | 7,692 | |
| ChAT-R | TTGGAAGCCATTTTGACTAT | 54.9 | |||
| Endogenous control | ACTB-F | ACCATGGATGATGATATCGC | 58.2 | 281 | 135 |
| ACTB-R | TCATTGTAGAAGGTGTGGTG | 54.4 | |||
| GABAergic/Glutaminergic neurons | GAD1-F | GTCGAGGACTCTGGACAGTA | 55.3 | 357 | 12,277 |
| GAD1-R | GGAAGCAGATCTCTAGCAAA | 54.9 | |||
| Serotonergic neurons | SLC6A4-F | GCCTTTTACATTGCTTCCTA | 54.8 | 447 | 2,251 |
| SLC6A4-R | CCAATTGGGTTTCAAGTAGA | 55.2 | |||
| Cholinergic neurons | ChAT-F | ACTGGGTGTCTGAGTACTGG | 55.0 | 451 | 7,692 |
| ChAT-R | TTGGAAGCCATTTTGACTAT | 54.9 | |||
| Dopaminergic neurons | TH-F | TCATCACCTGGTCACCAAGTT | 56.0 | 126 | 656 |
| GGTCGCCGTGCCTGTACT | 60.0 |
09-May-2012