| Products/Services Used | Details | Operation |
|---|---|---|
| Gene Synthesis | The 12 μL PCR reaction mix contained 50 ng DNA; 1.20 μL of 10X PCR buffer; 0.48 μL of 25 mM MgCl2 (Roche Applied Science, USA); 0.24 μL of 250 μM each of dCTP, dGTP, dTTP, and dATP (GenScript USA Inc. Piscataway, NJ); 0.06 μL of 10 μM M13-tailed forward primer; 0.30 μL of 10 μM reverse primer; 0.24 μL of 10 μM M13 primer fluorescently labeled with 6-FAM (6- carboxifluoresceine), Hex (hexachloro-fluoresceine), and NED [N (1-naphthyl) ethyletediamine] and 0.60 U Taq DNA Polymerase (New England Biolabs, Ipswich, MA, USA). PCR amplification was performed in 96-well plates on a Bio-Rad iCycler (Bio-Rad Laboratories Hercules, CA, USA) with an initial cycle of 94 °C for 3 min, 42 cycles of 1 min denaturing at 94 °C, 1 min annealing temperature 55/60 °C, 1 min extension at 72 °C, and the final extension at 72 °C for 10 min. The PCR products labeled with different fluorophores were mixed and detected on an ABI PRISM, 3130XI Genetic Analyzer (Applied Biosystems, Foster City, CA, USA). Amplified PCR products were size-separated by capillary electrophoresis on an Applied Biosystems fragment analyzer. Raw data files from fragment analyzers were imported into GeneMarker v1.91 (SoftGenetics LLC: www.softgenetics.com, 2004) to analyze amplicon sizes. | Get A Quote |
The preferential application of wheat flour in various end-use products is primarily driven by kernel texture (softness/hardness). Herein, a population of 268 F6recombinant inbred lines (RILs) was developed by crossing the wheat cultivarAlpowa (‘normal’ soft) (SKCS HI = 20.2) with a related ‘Super Soft’ line (BC2SS163) (HI = −1.2). A set of 19 polymorphic markers including 16 Kompetitive allele specific PCR (KASP) and three simple sequence repeat (SSR) markers were targeted to three genomic regions (4BS, 1BS and 5AL) and used to genotype the RILs and parents. Four QTLs were detected for kernel texture using composite interval mapping (CIM) including one major QTL on 4BS, and three minor QTL... More
