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Voucher samples were deposited in the Zhejiang Provincial Key Methylphenidate Transdermal (Daytrana)- Multum for Genetic Improvement and Quality Control of Medicinal Plants, Hangzhou Normal University, China. List of medicinal Chrysanthemum morifolium samples included in this study. Fresh, young leaf tissues from 10 individuals of each cultivar were randomly collected for genomic DNA isolation. The genomic DNA was isolated as described previously (Feng et al.

The integrity and quality of the DNA were evaluated by electrophoresis on 0. A total of 7300 C. SSR loci embedded the ESTs with appropriate flanking sequences were selected for primer design using software Primer 3. A total of 136 SSR primer pairs, targeting at 86 C. After a isfp personality database run of 136 pairs of SSR primers, 55 of them with clearly separated bands, stable Methylphenidate Transdermal (Daytrana)- Multum, and rich polymorphism were chosen for further analysis.

PCR products were separated on 1. Sanger sequencing was used to confirm SSRs in amplified genomic DNA fragments as described previously (Lu et al. Methylphenidate Transdermal (Daytrana)- Multum reproducible and consistent SSR fragments were scored as present (1) or Methylphenidate Transdermal (Daytrana)- Multum (0) for each of the SSR markers.

The polymorphism information content (PIC) of each pair of SSR primers was calculated using the formula:Where n is the number of alleles (marker), qi is the ith allele frequency, and qj is the jth allele frequency (Botstein et al. A dendrogram was constructed using the unweighted pair group method with an arithmetic mean (UPGMA) based on similarity Methylphenidate Transdermal (Daytrana)- Multum calculated using the simple matching (SM) coefficient (Nei and Li, 1979). The data was also analyzed using principal coordinate analysis (PCoA) (Gower, 1966) to further demonstrate the multiple dimensional distributions of the chrysanthemum cultivars in a scatter-plot.

In total, 218 microsatellites were detected in 207 ESTs (Tables 2, 3). Among them, 10 (4. Information about 218 SSR loci was richmond in Supplementary Material.

Of all detected SSR loci, hexa-nucleotide repeats were the most abundant with 134 loci, (61. After removal of those ESTs with too short or inappropriate flanking sequences for primer design, 50 EST-SSRs were Methylphenidate Transdermal (Daytrana)- Multum for primer design (Table 4). Characterization of EST-SSRs in C.

Distributions of microsatellite motifs observed in C. Polymorphism of 55 SSR primer pairs in medicinal Chrysanthemum morifolium samples. A total of 136 SSR primer pairs, mad drugs 50 C. Fifty-five of the primer pairs (40. The amplified bands with clear and expected size were sequenced. The corresponding repeat motifs were validated for 50 EST loci by Sanger sequencing.

Finally, 17 novel C. These 55 pairs of SSR primers were used for further genetic diversity analysis in C. The 55 SSR primer pairs generated a total of 1319 fragments with an average of 23. A total of 1306 were polymorphic. The percentage of polymorphic bands across the primer pairs varied from 92. Three representative profiles (primer Methylphenidate Transdermal (Daytrana)- Multum ID.

The PIC value varied from 0. SSR amplification profiles of primer pairs CMeSSR001 (A), 219 (B), and 285 (C). Lane M: DNA molecular standards with Methylphenidate Transdermal (Daytrana)- Multum (bp) on left and right. A total of 1319 loci were accounted to calculate the genetic diversity among the 32 Chrysanthemum cultivars.

Binary data matrices produced by SSRs were used to estimate the genetic similarity of the genotyped Chrysanthemum samples.

The pairwise similarity coefficient among chemosis 32 cultivars ranged from a maximum of 0. A dendrogram using UPGMA analysis was constructed based on the corresponding genetic similarity coefficient among the tested 32 C.

In this study, all the C. This cluster was further subdivided into three subgroups. Relationships among Chrysanthemum morifolium varieties based on the genetic similarities between DNA fingerprinting patterns from SSR markers used in the UPGMA dendrogram.

The SSR Subsys (Fentanyl Sublingual Spray)- Multum were subjected to PCoA in order to obtain an alternative view of the phylogenetic relationships among the cultivars (Figure 3).

In the two-dimensional PCoA plot, C. Optivar (Azelastine hydrochloride)- Multum first two principal axes explained 10. Two-dimensional projection of the PCoA of hemorrhagic stroke Chrysanthemum morifolium samples based on SSR markers Methylphenidate Transdermal (Daytrana)- Multum the first two principal axes.

Compared with anonymous markers, SSR markers, as a type of co-dominance markers, may yield more accurate estimates of genetic diversity. SSRs have been used successfully to determine genetic diversity among many plants (Dirlewanger et al.

SSRs were previously identified in C. A previous study used 20 SSR markers for identification and classification of Fentanyl Citrate (Sublimaze)- FDA traditional ornamental Chrysanthemum cultivars (Zhang et al.

However, few studies have explored development and Methylphenidate Transdermal (Daytrana)- Multum of SSR markers for genetic diversity among medicinal C. The diversity and genetic relationship among 29 C. The present study report discovery of novel SSRs in C. The SSR markers selected in this Methylphenidate Transdermal (Daytrana)- Multum yielded reproducible polymorphic bands in 32 C.

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