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The horticultural trait QTL originally detected by fine mapping each fractionated into two or more QTL at higher resolution.

P. pacificus and P. redivivus exudates were each fractionated by C18 solid phase extraction using a Varian Mega BE C18, which has a 5 mL void volume.

These two populations were each fractionated into three further groups CD34 positive, Sca1 negative (1 in Figure 3B; 4 in Figure 3C); CD34 negative, Sca1 negative (2 in Figure 3B; 5 in Figure 3C); or CD34 negative, Sca1 positive (3 in Figure 3B; 6 in Figure 3C and RNA was isolated from all six populations.

In our present study, the single QTL mapped for each trait in Brouwer and St. Clair (2004) each fractionated into multiple QTL after higher-resolution mapping, a phenomenon reported previously in other mapping studies (Chen and Tanksley 2004; Lecomte et al. 2004; Edwards and Mackay 2009; Studer and Doebley 2011; Johnson et al. 2012).

In addition, we investigated the thermal denaturation of each fractionated peak by DSF.

The expression status of NS in each fractionated cell type was confirmed by real-time RT PCR.

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All centrifugation steps and other operations were performed at 4°C. 100 µg of protein from each fraction was fractionated by SDS-polyacrylamide gel electrophoresis, blotted on to PVDF membrane using electrophoretic transfer cell (Bio-Rad) and probed with GLOI specific polyclonal antibody raised in mice (diluted 1∶500) [5].

Soon after, ACR was extracted using 80% EtOH (v/v; 2000 mL × 3, 48 hr each) and fractionated using n-hexane (2000 mL × 3, 48 hr each), to yield Ardisia crispa roots ethanolic hexane fraction (ACRH), as proposed previously [ 17].

Protein extracts (37 µg each) were fractionated on 17% SDS-PAGE and blotted on a nitrocellulose membrane (Whatman, Dassel, Germany).

Pooled CSF samples from CFS or nPTLS patients (total volume of 18 mL each), were fractionated using a 12.7×79.0 mm Seppro® IgY14 LC10 affinity LC column (Sigma, St Louis, MO) as previously described [18].

Protein extracts (20  μg each) were fractionated on proper concentration of SDS-PAGE and blotted on a nitrocellulose membrane (Whatman, Maidstone, UK; Cat. No.34000).

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