Thermo Fisher Scientific SuperScript IV Single Cell/Low Input cDNA PreAmp Kit, 48 reactions

Invitrogen™ SuperScript™ IV Single Cell/Low Input cDNA PreAmp Kit

The Invitrogen SuperScript IV Single Cell/Low Input cDNA PreAmp Kit is designed for efficient cDNA synthesis and amplification directly from intact single cells (1–1,000) or low amounts of total RNA (2 pg–10 ng).
It contains all required components to perform cell lysis, reverse transcription (RT), and PCR amplification in a convenient premixed format.
A combination of superior enzymes (SuperScript IV Reverse Transcriptase and Platinum SuperFi U DNA Polymerase) enables high yields, sensitivity, and accuracy.
Due to short reaction times and minimal hands-on time, this simple one-tube preamplification protocol can be completed in just over two hours.
The high-quality global cDNA obtained can be used for next-generation sequencing (NGS) or gene expression analysis by real-time PCR.

Features

• Superior sensitivity for detection of low-abundance targets (as low as single cell or 2 pg total RNA)
• High-quality cDNA providing full-length transcript information with uniform coverage
• Streamlined workflow with a simple one-tube protocol
• Short lysis and reverse transcription times for time savings
• Global preamplification compatible with downstream analysis by NGS or real-time PCR

Mechanism

The kit leverages the terminal deoxynucleotidyl transferase (TdT) activity of SuperScript IV Reverse Transcriptase.
Capturing oligonucleotides containing oligo(dT) and adapter sequences are used as primers for selective first-strand cDNA synthesis from poly(A)-containing RNAs.
When the reverse transcriptase reaches the 5′ end of the RNA template, TdT activity adds 1–3 extra nucleotides to the cDNA end, enabling template switching and ligation-free incorporation of adapter sequences at the 3′ end.
This process ensures high sensitivity and efficient capture of low-abundance targets.

Adapter sequences incorporated in both ends of the cDNA serve as primer-binding sites in the subsequent PCR amplification step, allowing global preamplification of full-length templates.
The included Platinum SuperFi U Preamplification Master Mix contains a high-fidelity DNA polymerase that ensures efficient amplification of long templates (up to 10 kb) with no PCR-induced bias or errors.

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