CapTrap-Seq: A platform-agnostic and quantitative approach for high-fidelity full-length RNA transcript sequencing
CapTrap-Seq: A platform-agnostic and quantitative approach for high-fidelity full-length RNA transcript sequencing
commercial samples obtained from Ambion: Ambion is a brand of life science research products and technologies, mainly focused on RNA products. It is owned by Thermo Fisher Scientific, a company that provides a variety of scientific research products and services. Therefore, \"total RNA commercial samples obtained from Ambion\" refers to RNA samples purchased from this brand for research purposes. SMARTer cDNA library preparation: SMARTer cDNA library preparation is a protocol for long-read sequencing that involves the addition of a poly(A) tail to RNA, followed by reverse transcription using oligo(dT) primers. This results in the synthesis of cDNA with a 5' end that contains a known sequence, which is used for further amplification and sequencing. The SMARTer protocol also incorporates a template switching oligo, which allows for the amplification of full-length cDNA molecules and increases the efficiency of library preparation. KAPA LTP Illumina Library Preparation kit: short read protocol to build libraries starting from full-length cDNA amplicons. The cDNA full-length libraries can be prepared using SMARTer or CapTrap full-length cDNA.  The cDNA samples were fragmented using Covaris under the following parameters: 10% Duty cycle, Intensity 5, 200 Cycles per burst, during 30 seconds in a total volume of 55 Î¼l. Illumina sequencing library was prepared using KAPA LTP Library Preparation kit Illumina and the manufacturer protocol was strictly followed. All samples were barcoded and multiplexed using the Illumina barcoding system (6 mers) and then sequenced in a Hi-Seq 2500 lane with HiSeq Sequencing v4 Chemistry.