A New Strategy for Identification of Highly Conserved microRNAs in Non-Model Insect, Spodoptera litura
Abstract
:1. Introduction
2. Results and Discussion
2.1. Amplification and Identification of sli-miR-14, sli-miR-2a and sli-bantam from S. litura
2.2. Cloning and Analysis of pre-miR-14 and pre-miR-2a from S. litura
2.3. Expression Patterns of sli-miR-14, sli-miR-2a and sli-bantam from S. litura
2.4. Discussion
3. Experimental Section
3.1. Sample Preparation
3.2. Stem-Loop RT-PCR Primers Designing
3.3. RNA Extraction and Reverse Transcription
3.4. PCR of sli-miR-14, sli-miR-2a and sli-bantam
3.5. Pre-miRNA Primers Designing
3.6. Genomic DNA Isolation and the Amplification of pre-miR-14 and pre-miR-2a from S. litura
3.7. Real-Time Quantitative PCR
3.8. Sequence and Data Analysis
4. Conclusion
Acknowledgments
- Conflict of Interest The authors declare no conflict of interest.
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RNA | linear equation | PCR efficiency | R2 |
---|---|---|---|
Sli-miR-14 | y = −3.3125x + 39.562 | 2.0039 | 0.9987 |
Sli-miR-2a | y = −3.2811x + 36.678 | 2.0173 | 0.9995 |
5S | y = −3.3915x + 33.219 | 1.9718 | 0.9945 |
miRNAs | Primer | Sequence (5′-3′) |
---|---|---|
Sli-miR-14 | RT | GTCGTATCCAGTGCAGGGTCCGAGGTATTCGCACTGGATACGACTAGGAG |
Forward | CGGGGCTCAGTCTTTTTCT | |
Reverse | GTGCAGGGTCCGAGGT | |
Sli-miR-2a | RT | GTCGTATCCAGTGCAGGGTCCGAGGTATTCGCACTGGATACGACGCTCAT |
Forward | GCCAGTATCACAGCCAGCT | |
Reverse | GTGCAGGGTCCGAGGT | |
Sli-bantam | RT | GTCGTATCCAGTGCAGGGTCCGAGGTATTCGCACTGGATACGACAATCAG |
Forward | GCCCCGTGAGATCATTTTG | |
Reverse | GTGCAGGGTCCGAGGT |
miRNAs | Primer | Sequence (5′-3′) |
---|---|---|
Sli-miR-14 | RT | GCACTTCAGTGTCGTGGTCAGTGACGGCAATTTGAAGTGCTAGGAG |
Forward | CGCACGACGCATCAGTCAGTCTTTTTCT | |
Reverse | GCACTTCAGTGTCGTGGTCAGTGACGGCAATT | |
Sli-miR-2a | RT | GCACTTCAGTGTCGTGGTCAGTGACGGCAATTTGAAGTGCGCTCAT |
Forward | CGACACACACCATCAGTATCACAGCCAGCT | |
Reverse | GCACTTCAGTGTCGTGGTCAGTGACGGCAATT | |
Sli-bantam | RT | GCACTTCAGTGTCGTGGTCAGTGACGGCAATTTGAAGTGCAATCAG |
Forward | CGCATCGTAGCATCGCTGAGATCATTTTG | |
Reverse | GCACTTCAGTGTCGTGGTCAGTGACGGCAATT |
© 2012 by the authors; licensee Molecular Diversity Preservation International, Basel, Switzerland. This article is an open-access article distributed under the terms and conditions of the Creative Commons Attribution license (http://creativecommons.org/licenses/by/3.0/).
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Gao, L.; Zuo, H.; Liu, K.; Li, H.; Zhong, G. A New Strategy for Identification of Highly Conserved microRNAs in Non-Model Insect, Spodoptera litura. Int. J. Mol. Sci. 2012, 13, 612-627. https://doi.org/10.3390/ijms13010612
Gao L, Zuo H, Liu K, Li H, Zhong G. A New Strategy for Identification of Highly Conserved microRNAs in Non-Model Insect, Spodoptera litura. International Journal of Molecular Sciences. 2012; 13(1):612-627. https://doi.org/10.3390/ijms13010612
Chicago/Turabian StyleGao, Lu, Hongliang Zuo, Keling Liu, Haiyi Li, and Guohua Zhong. 2012. "A New Strategy for Identification of Highly Conserved microRNAs in Non-Model Insect, Spodoptera litura" International Journal of Molecular Sciences 13, no. 1: 612-627. https://doi.org/10.3390/ijms13010612