A novel high-temperature One Step RT-PCR kit based on the combination of SunScript™ Reverse Transcriptase RNaseH- and high-quality Taq Polymerase for the reverse transcription and PCR amplification of specific RNA fragments in one convenient single step.
The SunScript™ One Step RT-PCR Kit is an easy and reliable system designed for fast, specific and sensitive end point RT-PCR reactions. This kit contains all the components needed to perform both reverse transcription and PCR amplification in a single tube by using gene specific primers in a “one step” reaction. This minimizes contaminations and constitutes a convenient system to process multiple samples.
100 ng of total RNA from HeLa cells were reverse transcribed and amplified with SunScript™ One Step RT-PCR Kit and specific primers for the indicated genes (GAPDH, HERC GUSB, CLTC and ACTIN).
The kit easily detects genes with vastly different expression levels.
100 ng of total RNA from HeLa cells were reverse transcribed and amplified with SunScript™ One Step RT-PCR Kit and specific primers for the GAPDH open reading frame (product length 305 bp).
The RT reaction can be performed at temperatures up to 85°C, making this the best choice for detecting difficult RNAs, with high degrees of secondary structure.
The indicated amounts of total RNA from HeLa cells were reverse transcribed and amplified with SunScript™ One Step RT-PCR Kit and specific primers for GAPDH cDNA (product length 305 bp).
The sensitivity of the kit is good enough to detect expressed genes from total RNA starting material in the order of picograms.
100 ng of total RNA from HeLa cells were reverse transcribed and amplified with SunScript™ One Step RT-PCR Kit and specific primers for the indicated genes (4 replicates are performed per gene).
The four independent replicates per gene show similar sensitivity and yield, making this kit the best choice for processing multiple samples with minimum deviations.
100 ng of total RNA from HeLa cells were reverse transcribed and amplified with SunScript™ RT-PCR Kit and four different specific primer pairs for a fragment of the TAFII100 cDNA with 75% GC content (product lengths 430-470 bp).
The RT-PCR protocol with sample denaturation was performed here and the cDNA synthesis step was performed at 60°C.
The four oligo pairs produced effective amplification bands despite the elevated GC content of the template RNA.
The SunScript™ One Step RT-PCR Kit has been tested in the reverse transcription / amplification of a 271 bp fragment of β-actin with a RNA dilution series.
The 2x Reaction Buffer contains a proprietary formulation which is optimized for both the reverse transcription and the PCR reaction and includes dNTPs, MgCl2 and stabilizers (to provide a final concentration of 250 nM of each dNTP and 3.5 mM MgCl2 in the reaction).
Each batch of SunScript™ One Step RT-PCR Kit is tested against predetermined specifications to ensure consistent product quality.
Shipping and Storage:
This product is shipped in dry ice. Upon receipt, it should be stored immediately at -20°C in a non-frost-free (constant temperature) freezer. If stored correctly, the product can be kept for at least six months after shipping without displaying any reduction in performance. For longer periods of time, store the kit at -80°C.