Description

ReverTra Ace™ qPCR RT Kit is an efficient and convenient kit to synthesize high quality cDNAs for real-time PCR. This kit contains the highly efficient reverse transcriptase "ReverTra Ace™" and a RT buffer optimized for the highly efficient synthesis of short-chain cDNAs suitable for real-time PCR. The protocol is simple, and the reaction can be completed in 15 min.

ReverTra Ace™ is a mutant-type M-MLV reverse transcriptase shows excellent efficiency.

ReverTra Ace™ qPCR RT Master Mix (Code No. FSQ-201) is a premix version of ReverTra Ace™ qPCR RT Kit. The master mix reagent (5x) contains the highly efficient reverse transcriptase "ReverTra Ace™" , primers and buffer optimized for highly efficient synthesis of short-chain cDNAs suitable for real-time PCR.

Features

• The optimized RT buffer and Primer Mix enable the highly efficient reverse transcription.
• The reaction can be completed in 15 min. The protocol does not contain an additional RNase H treatment step to remove residual RNA after reverse transcription (Patent Pending).
• Suitable for the detection of low-expressing mRNAs. Since the RT buffer is optimized for real-time PCR, the addition of 20% (v/v) of the synthesized cDNA solution to the PCR solution does not inhibit the PCR reaction.

Specification

Category	Item	Details
Storage condition	-	Store at -20ºC
ReverTra Ace™ qPCR RT Kit	5 x RT Buffer	400 µL
	Enzyme Mix	100 µL
	Primer Mix	100 µL
	Nuclease-free water	1000 µL × 2
ReverTra Ace™ qPCR RT Master Mix	5 x RT Master Mix	400 µL
	5 x RT Master Mix no RT-Control	40 µL
	Nuclease-free water	1000 µL × 2

Application

Reverse transcriptation for real-time PCR

Application Data

Example 1. Comparison of cDNA yields using various reverse transcription kits

cDNA was synthesized from human cell line (HeLa cell) total RNA with the ReverTra Ace™ qPCR RT kit. Subsequently, quantification of β-actin and polymerase ε mRNA were performed by real-time PCR with 100 ng cDNA, in conjunction with the SYBR® Green Realtime PCR Master Mix [Code No. QPK-201].

As shown in Fig. 1, the relative cDNA yields from the ReverTra Ace™ qPCR RT kit were greater than from the other commercial kits.

Comparison of relative cDNA concentrations

Fig. 1 Comparison of relative cDNA concentrations
The reaction temperature conditions of the reverse transcription are indicated under the figure.

Example 2. Detection of a rare expressing gene

Several 1-µL cDNA aliquots were prepared from 100 ng human cell line total RNA in a volume of 20 µL using various reverse transcription kits. Subsequently, a low expressing gene (TNF-α) was detected with real-time PCR kit (SYBR® Green Realtime PCR Master Mix [Code No. QPK-201]. The Ct values and success rates of the ReverTra Ace qPCR RT kit were greater than those from the other commercial kits (Fig. 2).

Fig. 2 Comparison of Ct values and success rates when detecting rare expressing genes

Documents

Download Manual Download SDS Request COA

ReverTra Ace qPCR RT Kit-Toyobo