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Detection of DNA Microsatellites Using Multiplex Polymerase Chain Reaction Aboard the International Space Station

Sophia Chen
Sophia Chen
, 
John Hatch
John Hatch
, 
Ashley Luck
Ashley Luck
, 
Nicole M. Nichols
Nicole M. Nichols
, 
Emily J. Gleason
Emily J. Gleason
, 
Kathryn Martin
Kathryn Martin
, 
Kevin D. Foley
Kevin D. Foley
, 
D. Scott Copeland
D. Scott Copeland
, 
Sebastian Kraves
Sebastian Kraves
 y 
Ezequiel Alvarez Saavedra
Ezequiel Alvarez Saavedra
   | 31 dic 2021
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Article Category: Research Note
Publicado en línea: 31 dic 2021
Páginas: 164 - 170
DOI: https://doi.org/10.2478/gsr-2021-0013
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© 2021 Sophia Chen et al., published by Sciendo
This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 3.0 License.

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).
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).
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).

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

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

Microsatellite markers targeted in this investigation.

Microsatellite marker Primer sequences Fluorescent label Expected size MSS (bp) Expected size MSI (bp)
NR-27 AACCATGCTTGCAAACCACTCGATAATACTAGCAATGACC FAM 90 87
NR-21 GAGTCGCTGGCACAGTTCTACTGGTCACTCGCGTTTACAA HEX 110 107
NR-24 GCTGAATTTTACCTCCTGACATTGTGCCATTGCATTCCAA NED 129 126
BAT-25 TACCAGGTGGCAAAGGGCATCTGCATTTTAACTATGGCTC FAM 152 148
BAT-26 CTGCGGTAATCAAGTTTTTAG AACCATTCAACATTTTTAACCC HEX 182 179
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