AQRNA-seq

AQRNA-seq enables quantitative mapping of small RNA populations to achieve absolute and linear quantification of microRNAs, tRNAs, RNA fragments, and ribonucleoside modifications.

Key Features:

• Bias Minimization: Reduces sequence-dependent biases occurring during capture, ligation, and amplification to improve representation of small RNA species.
• Quantitative Accuracy: Produces a direct, linear correlation between sequencing read count and RNA abundance validated with a 963-member microRNA reference library, RNA oligonucleotide standards of varying lengths, and northern blot.
• Versatility Across RNA Classes: Applies to challenging RNA classes such as bacterial tRNAs and reveals up to 80-fold variation in tRNA isoacceptor copy numbers, site-specific fragmentation patterns under stress, and quantitative maps of ribonucleoside modifications.
• Single-Experiment Mapping: Enables comprehensive mapping of small RNA landscapes in cells and tissues from a single experimental setup.
• Refined Library Preparation: Employs a library preparation process specifically modified to mitigate sequence-dependent biases.
• Computational Processing: Uses data mining algorithms and Python scripts integrating fastxtoolkit and BLAST for sequence processing and quantification.

Scientific Applications:

• Small RNA Quantification: Precise absolute and linear quantification of microRNAs, tRNAs, and other small RNAs.
• tRNA Dynamics and Stress Response: Analysis of tRNA isoacceptor abundance changes and site-specific fragmentation patterns under stress conditions.
• Epitranscriptomics: Mapping and quantification of ribonucleoside modifications across small RNA species.

Methodology:

Data processing uses Python scripts and data mining algorithms that integrate fastxtoolkit and BLAST to analyze AQRNA-seq libraries.