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| Status |
Public on Jan 31, 2015 |
| Title |
An improved protocol for small RNA library construction by using High-Definition adapters |
| Organism |
Medicago truncatula |
| Experiment type |
Non-coding RNA profiling by high throughput sequencing
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| Summary |
Most of small RNA library construction methods are based on RNA ligases, which prefer to join the molecules (small RNAs and adapters) that can anneal to each other and form a ligase favoured structure. Different platforms for next generation sequencing use different adapter sequences, causing the cloning bias. Adapters with degenerated nucleotides at the ligating ends (High Definition, HD adapters) were developed to reduce the cloning bias. However, above 90% of the cloning products is adapter dimer when the current available commercial kits and their corresponding protocols are used. Here we adopted and further improved a method demonstrated in a publically available patent (http://www.google.com/patents/WO2011056866A2?cl=en). Using the improved method, we constructed the small RNA libraries by using the total RNA of Medicago truncatula leaf tissue. The adapter dimer was significantly reduced. The small RNA sequences were also analysed.
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| Overall design |
The small RNA libraries of medicago truncatula leaves were constructed by using an improved protocol where high-definition (HD) adapters were used.
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| Contributor(s) |
Xu P, Billmeier M, Mohorianu I, Green D, Fraser WD, Dalmay T |
| Citation |
https://doi.org/10.1515/mngs-2015-0001
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| |
| Submission date |
Jul 11, 2014 |
| Last update date |
May 15, 2019 |
| Contact name |
Irina Mohorianu |
| E-mail(s) |
[email protected]
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| Organization name |
University of Oxford
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| Department |
Medical Sciences Division
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| Lab |
Oxford Vaccine Group
|
| Street address |
Headington
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| City |
Oxford |
| ZIP/Postal code |
OX3 7LE |
| Country |
United Kingdom |
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| Platforms (1) |
| GPL17491 |
Illumina HiSeq 2000 (Medicago truncatula) |
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| Samples (2) |
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| Relations |
| BioProject |
PRJNA255087 |
| SRA |
SRP044263 |
