Table 1.
TNR contraction data in SVG-A cells
| 1 | 2 | 3 | 4 | 5 | 6 | 7 | 8 | 9 | 10 | |
|---|---|---|---|---|---|---|---|---|---|---|
| Repeat sequence | Plasmid name | Vector backbone | SVG-A contractions observed (His+ Ura+ colonies) | Plasmid population size (His+ colonies × dilution) | N | SVG-A contraction frequency% (±SEM%)a | Background contraction frequency% (±SEM%) | Corrected contraction frequency | P-value: SVG-A versus Background | |
| A | (C,A,G)33= 0 rpts | pBL200 | pBL67 | 0 | 6000 | 3 | 0 (±0) | 0 (±0) | 0 | N/A |
| B | (CAG)15+18 | pBL213 | pBL67 | 24 | 392 400 | 6 | 0.0079 (±0.0029) | 0.0077 (±0.0086) | 0.0002 | 0.91 |
| C | (CAG)25+8 | pBL176 | pBL67 | 2923 | 789 000 | 5 | 0.34 (±0.09) | 0.16 (±0.02) | 0.19 | 0.0004 |
| D | (CTG)25+8 | pBL207 | pBL179 | 10 | 29 800 | 3 | 0.04 (±0.02) | 0.02 (0.04) | 0.02 | 0.56 |
| E | (CAG)30+3 | pBL227 | pBL67 | 204 | 70 800 | 6 | 0.82 (±0.28) | 0.43 (±0.12) | 0.39 | 0.18 |
| F | (CAG)33 | pBL217 | pBL67 | 622 | 37 600 | 4 | 2.3 (±0.4) | 0.31 (±0.11) | 1.9 | 0.01 |
| G | (CTG)25+8 | pBL199 | pBL67 | 8 | 22 200 | 2 | 0.034 (±0.006)a | 0.015 (±0.009) | 0.019 | 0.10 |
| H | (CTG)50 | pBL175 | pBL67 | 37 | 12 800 | 3 | 0.30 (±0.06) | 0.055 (±0.014) | 0.24 | 0.002 |
| I | (CAG)25+8 | pBL250 | pBL245 | 49 | 34 600 | 4 | 0.17 (±0.03) | 0.003 (±0.002) | 0.17 | 0.006 |
| J | (TAG)25+8 | pBL246 | pBL245 | 4 | 6000 | 2 | 0.067 (±0.009)a | 0.013 (±0.013) | 0.054 | 0.05 |
| K | (CTA)50 | pBL216 | pBL67 | 173 | 274 000 | 5 | 0.09 (±0.05) | 0.06 (±0.01) | 0.03 | 0.89 |
| L | (CTG)6(ATG)2(CTG)25 | pBL248 | pBL245 | 576 | 60 000 | 6 | 0.87 (±0.07) | 0.051 (±0.004) | 0.82 | 0.0001 |
| M | (CTG)33 | pBL249 | pBL245 | 50 | 12 800 | 3 | 0.39 (±0.01) | 0.12 (±0.02) | 0.27 | 0.05 |
| N | (CAG)33 | pBL247 | pBL245 | 2421 | 118 400 | 3 | 2.0 (±0.5) | 0.14 (±0.01) | 1.9 | 0.04 |
Column 1 lists the trinucleotide repeat sequence comprising the template for lagging strand synthesis (as defined in Materials and Methods). Column 2 gives each plasmid's name for reference. Column 3 indicates the vector backbone in which each TNR sequence was cloned. For (CAG)25+8 and (CAG)33, the repeats were cloned in both vector backbones—the pBL67 derivatives (pBL176 and pBL217) were used in the threshold study while the pBL245 vectors (pBL250 and pBL247) were used in the hairpin and orientation dependence studies. The pBL67 derivative, pBL217, was also used as the (CAG) 33-new ori vector in the orientation dependence study. Column 4 indicates the total number of contractions observed in triplicate transfections in SVG-A cells (the number of colonies counted on SC-His-Ura plates). Column 5 lists the total number of plasmids in the population (the number of colonies counted on SC-His plates multiplied by the appropriate dilution factor). Column 6 lists the number of samples recovered after transfection. Column 7 lists the cntraction frequency observed for plasmids replicated in SVG-A cells (%) ±1 SEM (%). The error for samples with two replicates (marked ‘a’) is listed as the absolute difference between the two observed values. Column 8 lists the background contraction frequency (%) ±1 SEM (%), as described in Materials and Methods. Column 9 lists the corrected contraction frequency, obtained by subtracting the background (column 8) from the contraction frequency observed in SVG-A cells (column 7). The corrected contraction frequency is used to produce the graphs shown in the figures. Column 10 lists the P-value for each sequence versus the background as determined by two-tailed student's t-test.
aError shown is the range between the two values.