Three different SCAR markers were amplified: two specific to strain Ach 1-1 (189 bp and 387 bp) and one specific to strain 1113-5 (431 bp).
Seven breast cancer genetic markers were amplified by means of the MLPA reaction, which allows for multiplex amplification of multiple targets with a single primer pair.
GW2-specific CAPS markers were amplified using the primers GW2-Hinf1-F and GW2-Hinf1-R.
All ctDNA markers were amplified with NEB Taq using the buffer supplied by the manufacturer (NEB co., Japan).
SSRs, and indel and dCAPS markers were amplified using standard PCR protocols and run in 3% agarose gel with ethidium bromide.
All markers were amplified using the same standard PCR protocol.
HIS3 and TRP1 markers were amplified from plasmids pRS413 and pGBKT7 respectively.
Comparisons with the GenBank database confirmed that the correct markers were amplified, with the highest sequence similarity to other Montipora sequences in the database.
Microsatellite markers were amplified using the Multiplex PCR Kit (Qiagen) and fragment size analyses were determined on an ABI 3730 capillary sequencer (Applied Biosystems) and analyzed with the GeneMapper 4.0 software (Applied Biosystems).
In Uganda where transmission intensity is very high, selected regions of MSP1 and MSP2 and 4 microsatellite markers were amplified using PCR and characterized based on sequence and size polymorphisms identified by gel electrophoresis [16].
Microsatellite markers were amplified using 1 µl DNA (∼20 ng), 0.2 µM each primer, 50 µM dNTPs, 1x polymerase chain reaction (PCR) buffer (Roche; 10 mM Tris HCl, pH 8.3, 1.5 mM MgCl2, 50 mM KCl) and 0.5 U Taq polymerase in a final reaction volume of 20 µl.
