Table S1
Amplification reactions run on the ISS.
| Reaction 1 | DNA | Primers |
|---|---|---|
| 1 | MSS | Pentaplex: NR-27, NR-21, NR-24, BAT-25, BAT-26 |
| 2 | MSI | Pentaplex: NR-27, NR-21, NR-24, BAT-25, BAT-26 |
| 3 | MSS | NR-27 |
| 4 | MSI | NR-27 |
| 5 | MSS | NR-21 |
| 6 | MSI | NR-21 |
| 7 | MSS | NR-24 |
| 8 | MSI | NR-24 |
| Reaction 2 | DNA | Primers |
|---|---|---|
| 1 | MSS | Pentaplex: NR-27, NR-21, NR-24, BAT-25, BAT-26 |
| 2 | MSI | Pentaplex: NR-27, NR-21, NR-24, BAT-25, BAT-26 |
| 3 | MSS | BAT-25 |
| 4 | MSI | BAT-25 |
| 5 | MSS | BAT-26 |
| 6 | MSI | BAT-26 |
| 7 | MSS | Negative control |
| 8 | MSI | Pentaplex: NR-27, NR-21, NR-24, BAT-25, BAT-26 |
[i] Single reactions for all five primer sets and pentaplex reactions were run on both DNA templates (MSS and MSI). A no-primer reaction was run as a negative control.
[ii] ISS, International Space Station; MSI, microsatellite instability; MSS, microsatellite-stable.
Table S2
Microsatellite markers targeted in this investigation.
| Microsatellite marker | Primer sequences | Fluorescent label | Expected size MSS (bp) | Expected size MSI (bp) |
|---|---|---|---|---|
| NR-27 | AACCATGCTTGCAAACCACT CGATAATACTAGCAATGACC | FAM | 90 | 87 |
| NR-21 | GAGTCGCTGGCACAGTTCTA CTGGTCACTCGCGTTTACAA | HEX | 110 | 107 |
| NR-24 | GCTGAATTTTACCTCCTGAC ATTGTGCCATTGCATTCCAA | NED | 129 | 126 |
| BAT-25 | TACCAGGTGGCAAAGGGCA TCTGCATTTTAACTATGGCTC | FAM | 152 | 148 |
| BAT-26 | CTGCGGTAATCAAGTTTTTAG AACCATTCAACATTTTTAACCC | HEX | 182 | 179 |
[i] Primer sequences for and expected sizes in MSS and MSI cell lines. Reverse primers were fluorescently labeled to facilitate separation by capillary electrophoresis.
[ii] MSI, microsatellite instability; MSS, microsatellite-stable.
Figure 1
Experimental approach and gel electrophoresis results. (A) PCR samples were prepared on the ground and frozen at −80°C prior to launch to ISS. PCR took place aboard the ISS and amplified samples were frozen at −80°C for return to Earth. Electrophoresis was carried out on the ground. (B) Gel electrophoresis image of PCR products. NR-27, NR-21, NIR-24, BAT-25, and BAT-26 are the five microsatellites. MSS and MSI indicate the source of template DNA. Samples were amplified either on the ground (G) or in space (S).
Figure 2
Determination of microsatellite sizes of pentaplex reactions by capillary electrophoresis. Capillary electrophoresis traces of pentaplex amplification reactions from either MSI or MSS samples performed either on the ground or in space show amplification of all five target sequences (NR-27, FAM channel, blue; NR-21, PET channel, red; NR-24, VIC channel, green; BAT-25, FAM channel, blue; BAT-26, PET channel, red).
Table S3
Amplicon size comparison for single and pentaplex PCR.
| NR-27 | NR-21 | NR-24 | BAT-25 | BAT-26 | ||||||
|---|---|---|---|---|---|---|---|---|---|---|
| Penta | Single | Penta | Single | Penta | Single | Penta | Single | Penta | Single | |
| Ground MSI | 74.7 | 73.6 | 102.6 | 102.8 | 115.8 | 115.9 | 141.7 | 141.7 | 168.4 | 168.9 |
| Space MSI | 73.9 | 73.9 | 102.1 | 102.1 | 115.5 | 115.8 | 141.6 | 141.6 | 168.3 | 168.2 |
| Ground MSS | 84.6 | 84.6 | 107.2 | 108.0 | 122.4 | 122.4 | 145.5 | 145.4 | 180.7 | 180.6 |
| Space MSS | 84.5 | 84.5 | 107.2 | 107.2 | 122.4 | 122.3 | 145.4 | 144.5 | 180.6 | 180.5 |
[i] Quantification of allele size determined for each amplicon in pentaplex reactions yields allele sizes consistent with those determined in single reactions, regardless of sample type (MSI vs. MSS) or PCR conditions (ground vs. space).
[ii] MSI, microsatellite instability; MSS, microsatellite-stable; PCR, polymerase chain reaction.