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

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 the 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) applications or for gene expression analysis by real-time PCR.

Features of the SuperScript IV Single Cell/Low Input cDNA PreAmp Kit include:
• Superior sensitivity for easy detection of low abundant targets (as low as single cell or 2 pg total RNA)
• High quality cDNA for full-length transcript information with uniform coverage
• Streamlined workflow with 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

The SuperScript IV Single Cell/Low Input cDNA PreAmp Kit leverages the terminal deoxynucleotidyl transferase (TdT) activity of the SuperScript IV Reverse Transcriptase. Capturing oligonucleotide containing oligo(dT) and adapter sequences is used as primer 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 sequence in the 3‘ end of the resulting cDNA. The high sensitivity and template switching efficiency of SuperScript IV Reverse Transcriptase allow the capture of even 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 the full-length templates. The included Platinum SuperFi U Preamplification Master Mix contains a superior high-fidelity DNA polymerase that ensures efficient amplification of long templates (up to 10 kb) with no PCR-induced bias or errors.