AcuGenix™ First-Strand cDNA Synthesis Kit (for Single-Cell 3′ RNA-Seq)

AcuGenix™ First-Strand cDNA Synthesis Kit (for Single-cell 3’RNA sequencing)

Template conversion technology empowers the synthesis of full-length cDNA for high-fidelity single-cell transcriptome research

This kit is specifically developed for single-cell 3' RNA sequencing scenarios of mammalian cells or eukaryotic cells without cell walls. Through engineered reverse transcriptase and an optimized template conversion process, it can efficiently synthesize full-length first-strand cDNA that retains the 5 'end sequence information of mRNA. The product carries known fixed sequences at both ends and can be directly used for downstream sequencing library construction.

1. Template conversion for full-length cDNA synthesis:
Engineered reverse transcriptase initiates reverse transcription by binding to the poly(A) tail of mRNA through oligo(dT) primers. After reaching the 5 'end of the mRNA, its terminal transferase activity adds three cytosine (CCC) tails to the 3' end of the newly synthesized cDNA. This poly(C) tail can serve as an anchor site and bind to the pre-added template conversion oligonucleotide (TSO) - the 3 'end of the TSO carries three complementary guanines (GGGS), triggering the template conversion effect. The reverse transcriptase continues to extend to the TSO end, ultimately obtaining a full-length cDNA with standardized sequences at both ends. Avoid the 5 'end preference of traditional reverse transcription.

2. Outstanding performance of engineered reverse transcriptase:
Compared with wild-type M-MLV reverse transcriptase, the engineered enzyme used in this kit has significantly improved thermal stability, better reverse transcription efficiency, and tolerance to high concentrations of DTT. It can maintain robust performance under various reaction conditions.

3. Wide template input range compatibility
It is compatible with total RNA containing poly(A) tails ranging from 10 pg to 1 μg, and simultaneously supports the requirements of single-cell and low starting dose conventional RNA experiments.

4. Seamless connection of downstream processes
The two-end sequences of the obtained full-length cDNA are standardized and can be directly used in standard processes such as cDNA amplification, enzymatic digestion and fragmentation, and sequencing library construction, without the need for additional modification steps.

High-throughput single-cell 3’RNA sequencing (scRNA-seq).

2. Transcriptome analysis of rare cell populations (such as circulating tumor cells, antigen-specific immune cells).

3. Low initial transcriptome study of trace eukaryotic cell samples.

4. Construction of full-length cDNA libraries required for the discovery of transcript isomers.