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©2010 Baishideng Publishing Group Co.
World J Stem Cells. Dec 26, 2010; 2(6): 127-140
Published online Dec 26, 2010. doi: 10.4252/wjsc.v2.i6.127
Published online Dec 26, 2010. doi: 10.4252/wjsc.v2.i6.127
Table 2 Comparative targeting efficiencies of R1 and HM-1 mouse embryonic stem cells
| Parameters | Cell line (passage number) | |||||||||
| R1 (p12) | HM-1 (p21) | |||||||||
| Amaxa | BioRad | ∑ | Amaxa | BioRad | ∑ | |||||
| Electroporation program | A231 | A241 | A301 | A231 | A241 | A301 | ||||
| Cell number | 3 × 106 | 3 × 107 | 3 × 106 | 3 × 107 | ||||||
| DNA concentration (μg) | 5 | 10 | 5 | 10 | ||||||
| Number of selected clones | 120 | 192 | 168 | 356 | 836 | 168 | 168 | 24 | 384 | 744 |
| Positive clones after 1st PCR screen | 3 | 10 | 11 | 15 | 39 | 5 | 4 | 0 | 15 | 24 |
| Percentage (%) | 2.5 | 5.2 | 6.5 | 4.2 | 4.7 | 2.9 | 2.4 | 0 | 3.9 | 3.2 |
| Positive clones after 2nd PCR screen | 3 | 10 | 11 | 15 | 39 | 5 | 4 | 0 | 15 | 24 |
| Homologous recombination confirmed by Southern blot analysis | ||||||||||
| Clone number | 3 | 9 | 10 | 7 | 29 | 4 | 4 | 0 | 5 | 14 |
| Efficiency (%) | 2.5 | 4.7 | 6 | 2 | 3.5 | 2.4 | 2.4 | 0 | 1.3 | 1.9 |
| Transfection efficiency | ||||||||||
| Transient | No data on Amaxa system | 91% ± 6.1% | No data on Amaxa system | 75% ± 4.2% | ||||||
| Stable | No data on Amaxa system | 34% ± 2.9% | No data on Amaxa system | 25% ± 5.3% | ||||||
- Citation: Mamo S, Kobolak J, Borbíró I, Bíró T, Bock I, Dinnyes A. Gene targeting and Calcium handling efficiencies in mouse embryonic stem cell lines. World J Stem Cells 2010; 2(6): 127-140
- URL: https://www.wjgnet.com/1948-0210/full/v2/i6/127.htm
- DOI: https://dx.doi.org/10.4252/wjsc.v2.i6.127