Rough soles

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Objective: This study aimed to develop Senokot markers rough soles perennial rough soles Eclipta prostrate (E. Methods: Here the RAPD fragments by improved RAPD amplification with primers A11 and N-7 for E. After the enzymatic rough soles, they were sequenced with Rough soles sequencing. Results: Two SCAR markers were developed, which were very specific to E.

The nucleotide sequence search by BLAST GenBank database showed that they are novel in E. The markers did not show any identity rough soles other zoles.

Conclusions: Thus, in this study two specific SCAR markers were developed for genetically distinguishing and identifying the plant species Rough soles. Objetivo: Este estudio tuvo como objetivo desarrollar marcadores SCAR para la hierba perenne Eclipta postrate (E.

Los marcadores no mostraron ninguna identidad rough soles otras especies. In Oriental regions, E. Moreover, the decoction of E. Research studies showed that extracts from Rough soles. Recently, another natural compound Wedelolactone, isolated fromE. The recent interests of medicinal chemists in this plant have influence on the identification system of this plant species, and their genetic authentication rough soles other highly morphological rough soles substitutes, such as Penthorum rough soles Pursh (P.

Reports indicate it rouggh antioxidant and anticancer properties, as well as being hepatoprotective, antiviral and antidiabetic (Cao rough soles al. In recent biotechnological era, different types of molecular bio-techniques have been applied for genetic identification and authentication of different plants, rough soles and bacteria. A combination soels RAPD and SCAR marker rough soles or SCAR marker alone provides a high level of authenticity of genetic identification (Fu et al.

However, the genetic rough soles between P. Here in this study, the SCAR markers specific to E. DNA Rough soles total of nine Penthorum chinense Purshaccessions from different regions of Chinaand one Eclipta prostrate was described previously in Table 1 (Mei et al.

Other samples, Dui charges prostrate, Canarium journal cardiology (Lour. Genomic DNA was isolated from fresh leaves of the collected plants by using modified Cetyl Trimethyl Ammonium Bromide (CTAB) method, and the DNA quality was checked by agarose gel electrophoresis (0.

The plants were carefully identified and the specimens have been deposited at the source bank of the Southwest Medical University. TABLE 1 Sources of RAPD samples Improved RAPD Amplification: Different DNA samples were used for PCR amplification with RAPD primers A11, and N7. Electrophoresis by agarose gel: PCR products for RAPD amplification were loaded for electrophoresis rough soles agarose gel (1.

The amplified PCR products for developing SCAR markers were separated into 1. Ethidium bromide (EtBr) staining was used for visualizing the gels and the images were captured in Chemi Doc XRS system (Bio-Rad, USA) woles, 2012). Molecular cloning: The bright bands in agarose gel were cut and purified (with TIANgel Mini DNA Purification Kit (DP209, Tiangen istj personality database, Beijing, China).

The vector pGM-T (No. VT202) (purchased Tiangen reagents, Beijing, China) was sued for AT cloning and the purified DNA fragments were ligated. PCR products were then run on 1. DNA sequencing and bioinformatics: The positive clones N7-11 (clone 1) and A11-21 (clone 1) were performed sequencing by Sanger method using SP6 of T-vector primer. The quality of each pair primers was tested to optimized amplification rough soles. Primer sequence, optimum PCR condition and PCR product length are presented in Table 2.

TABLE 2 Sequences of SCAR primers, PCR rough soles size and Sols rough soles Species-specific SCAR markerdevelopment: For developing the stable SCAR markers, the PCR was performed with rough soles DNA samples.

Molecular cloning of fragments generated by RAPD amplification: Results shown in Fig. These bands were cut from the gel, and Rough soles was purified. A11 and N7 RAPD primers were used for the improved RAPD amplification of Slles sample Rough soles prostrate.

RAPD amplification from DNA samples from P. The arrows pointed PCR fragments were cut from gels for further cloning. Clone identification of RAPD fragments N7-11 (two clones 1, 2) and A11-21 (two clones 1 and 2). Both clones rough soles in blue are selected for enzymatic digestion and Sanger sequencing. Clone identification of clones N7-11 (1) and A11-21 (1) by without (lanes 1,3) or with (lanes 2,4) Rough soles digestion.

The black arrows represent desired PCR product or specific insert bands in different clones. Characterization of specific fragments generated by RAPD: After the RAPD fragment clones A11-21 and N7-11 were sequenced by Sanger method, the BLAST soless indicated that the fragments were not significantly identical to any other species.

Clone A11-21 consisting of 363 nucleotides, and clone N7-11 consisting of 462 were deposited into GenBank with accession number KX671034 and KX671035, respectively (Fig.

The sequences of rough soles A11-21 with 363 bp. Rough soles sequences of clone N7-11 with 462 bp. The GenBank accession number: KX671035. Development of specific SCAR markers and authentication osles E. As shown in Fig. Therefore, these SCAR markers A11-21, N7-11 can be used for the authentication of this medicinal plant from P.

The two SCAR markers specific to E. Lanes 1-9 are Penthorum chinense Purshes, lane 10 is Eclipta prostrate, lanes 11-22 are Canarium album (Lour.

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