MicroRNAs in Development: Methods and Protocols

MicroRNAs in Development: Methods and Protocols (Methods in Molecular Biology)
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  • Inspired by Your Shopping History
  • Detection of mRNA and microRNA Expression in Basal Chordates, Amphioxus and Ascidians
  • MicroRNA sequencing - Wikipedia
  • Supplementary files
  • Introduction

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    Sign In Register Help Cart. M values calculated by geNorm represent the stability of each gene with smallest values indicative of highest expression stability. NormFinder is another acknowledged algorithm used for determining the stability of reference genes with smaller values representing higher expression stability. For genes where expression was significantly different over time, pair wise differences between time points was assessed with Tukey's HSD tests.

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    From miRBase Following our criteria described above, 1, candidate miRNA loci were identified in the D. After the secondary structure analysis by mFold, a total of 75 D. After several attempts to amplify RNU1A and U6 using sequence specific primers, only the U6 gene could be successfully amplified. This is not surprising since U6 is one of the most highly conserved spliceosomal RNAs [21]. U6 gene had the smallest M value 0.

    Detection of mRNA and microRNA Expression in Basal Chordates, Amphioxus and Ascidians

    Out of the 14 miRNAs selected for validation, 8 mir-8, mir-9, mir, mir, mir, mir, mir and mir were successfully amplified. This suggests that our computational prediction method is an efficient way to discover conserved miRNAs in D. Since only one pair of primers was tested for each miRNA, additional primers might need to be designed for further validation of the remaining 6 miRNAs mir-1, mir, mir, mir, mir and mir Because mir expression could only be detected at day 12, it was excluded from the expression stability test and expression changes test. With exception of only two miRNAs mir and mir all miRNAs changed in expression during the three time points monitored, with only one miRNA mir decreasing in expression at days 8 and 12 Figure 1.

    The significant up-regulation of mir-8 at day 12 and of mir-9 at day 8 coincides with dramatic morphological changes including molting, somatic growth, brood chamber development and egg development in D. Thus, we propose that mir-8 and mir-9 might also play important roles in D. The roles of mir and mir in D. Interestingly, mir and mir are located within a 1 kb region and transcribe the same pri-miRNA, but had opposite expression patterns.

    MicroRNA sequencing - Wikipedia

    This uncoordinated expression profile has also been identified in Drosophila [30] and provides evidence of post-translational regulation in these clustered miRNAs. In this research, we predicted 75 conserved D. Significant changes in the expression of mir-8, mir-9, mir, mir and mir were observed, suggesting they play an important role during Daphnia development.

    Supplementary files

    Since the discovery of microRNAs, developmental biologists have striven to understand the role of miRNAs in development and disease. MicroRNAs in. Read MicroRNAs in Development: Methods and Protocols: (Methods in Molecular Biology) book reviews & author details and more at giuliettasprint.konfer.eu

    As a next step, a specific designed D. This study is the first to report expression of miRNAs on D. The authors acknowledge Dr. John Colbourne's lab at Indiana University for providing D.

    Methods and Protocols

    Matthew Hale for their valuable comments. Predicted D. Biological functions associated with the differentially expressed miRNAs during D. Wrote the paper: SC. Drafted the manuscript: SC. Browse Subject Areas? Click through the PLOS taxonomy to find articles in your field. Abstract Daphnia pulex , the crustacean with the first sequenced genome, is an important organism that has been widely used in ecological and toxicological research.

    Introduction Daphnia pulex is an ecologically important organism in freshwater ecosystems that has been studied for decades [1] and is the first crustacean for which a full genome sequence is available. Materials and Methods In silico prediction of D. In vivo validation of D.

    Introduction

    This technique is dependent on hydrogel particles that contain custom selected probes against target miRNAs. Varallyay, E. Incubate in a fume hood at room temperature overnight. Biochem Biophys Res Commun , — Data analysis can also be relatively challenging, especially for a labs that may not have a resident bioinformatician. Hou, Y. Angermueller, C.

    Expression of D. Prime members enjoy FREE Delivery on millions of eligible domestic and international items, in addition to exclusive access to movies, TV shows, and more.

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