Unknown,Transcriptomics,Genomics,Proteomics

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An improved protocol for small RNA library construction by using High-Definition adapters

ABSTRACT: 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. The small RNA libraries of medicago truncatula leaves were constructed by using an improved protocol where high-definition (HD) adapters were used.

ORGANISM(S): Medicago truncatula

SUBMITTER: Irina Mohorianu

PROVIDER: E-GEOD-59330 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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An improved protocol for small RNA library construction by using High-Definition adapters

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