Explore how RNA-seq adaptors play a critical role in RNA sequencing:
Fragmentation and Reverse Transcription:
During library preparation, RNA is often fragmented to generate short cDNA fragments that can be easily sequenced. Adaptors are ligated to the fragmented RNA molecules, providing the necessary priming sites for reverse transcription.
Addition of Unique Identifiers:
Adaptors often include unique molecular identifiers (UMIs) or barcodes that tag individual RNA molecules. This helps distinguish between identical sequences derived from different RNA molecules, providing accurate quantification and reducing PCR amplification biases.
Sequencing Platform Compatibility:
Adaptors are designed to be compatible with the specific sequencing platform being used (e.g., Illumina, Oxford Nanopore, PacBio). They contain sequences that allow for proper binding to the flow cells or nanopores and facilitate the sequencing process.
Multiplexing:
Adaptors enable multiplexing, allowing multiple samples to be sequenced together in a single run. Unique barcodes within the adaptors help in demultiplexing the sequenced data and assigning reads to their respective samples.
