Invitrogen™ Oligonucleótidos de transcripción inversa con cambio de plantilla

Descripción

Este conjunto de oligonucleótidos está diseñado para la reacción de transcripción inversa con cambio de plantilla y la posterior preamplificación del ADNc obtenido. El oligonucleótido de cambio de plantilla (TSO) y el oligonucleótido de captura (dT) se utilizan durante la reacción de síntesis de ADNc para incorporar secuencias adaptadoras en ambos extremos del ADNc de longitud completa. El cebador de preamplificación es complementario a la secuencia del adaptador y permite la amplificación por PCR del ADNc obtenido.

Template Switching Oligonucleotide (TSO) and Capturing Oligo (dT) Primer are designed for use in template switching reverse transcription reaction. Capturing oligonucleotide containing oligo(dT) and adapter sequences is used as a primer for selective first-strand cDNA synthesis from poly(A)-containing RNAs. When the reverse transcriptase reaches the 5ʹ end of the RNA template, terminal deoxynucleotidyl transferase (TdT) activity of reverse transcriptase adds 1–3 extra nucleotides to the cDNA end, enabling hybridization of the template switching oligo and ligation-free incorporation of adapter sequence to the 3ʹ end of the resulting cDNA. Both oligos are supplied as ready-to-use 20X concentrated solutions. For maximal template switching efficiency, we recommend using Template Switching Oligo and Capturing oligo (dT) Primer with Invitrogen SuperScript IV Template Switching RT Master Mix .

Adapter sequences incorporated at both ends of the cDNA may serve as primer-binding sites in the subsequent PCR amplification step, allowing global preamplification of full-length templates. For PCR, use Preamplification Primer which is complementary to TSO and Capturing Oligo (dT) adaptor sequence and is supplied as ready-to-use 100X concentrated solution.

The TSO, Capturing Oligo (dT) and Preamplification primers are also available as part of complete Invitrogen SuperScript IV Single Cell/Low Input cDNA PreAmp Kit (Cat. No. 11752048) .

Applications
• cDNA synthesis
• cDNA preamplification
• RNA-seq
• RACE