Protocol: cDNA Synthesis and qRT-PCR using the BioRad MyiQ

Hy forum fella,
This is krusty... new for you forum members, new for me. Useful tool! (I hope)
Is there anyone with an indication of the expected cDNA yield after a cycle of iScript cDNA synth. kit (BioRad)? I know it depends on RNA quality and quantity, but I only wanted to know if it is correct to expect a 100% transcription efficiency in the case of a "perfect" RNA sample, or if this percentage is lower. And, in the case of lower quality and low-concentrated RNA (c.a. 15 ng RNA), is it likely to expect very low efficiency? (e.g. 10% or lower)
Any help is appreciated, any reference as well.
Thanks in advance
Bests
K

iScript 5x Reaction Buffer (blue tubes, -20°C common freezer)

iScript Reverse Transcriptase (yellow tubes, -20°C common freezer)

iScript™ Advanced cDNA Synthesis Kit for RT-qPCR

For 2-step RT-qPCR, Bioline has developed the SensiFAST™ cDNA Synthesis Kit to complement the SensiFAST Probe and SYBR® Kits. The unique combination of our high-performance reverse transcriptase and optimized TransAmp™ buffer system delivers excellent reverse transcription efficiency across a very broad range of input RNA concentrations.

iScript™ Reverse Transcription Supermix for RT-qPCR

The SensiFAST™ Genotyping Kit has been developed for high-performance dual-labelled probe assays that deliver fast, precise and highly reproducible characterization of sequence variants, including loci with type IV SNPs. The optimized buffer system promotes clear allelic discrimination and outstanding allele clustering.

1. Thaw RNA samples, spec using NanoDrop (8th floor Stearns). Thaw iScript 5x reaction buffer.

Kuperwasser Lab | cDNA synthesis and q-PCR (APS)

The product featured in the video, Bio-Rad’s , is a simple and sensitive first-strand cDNA synthesis kit for gene expression analysis. The supermix reduces chances of error with a one-tube setup and cuts bench time in half.

Biorad iscript cdna synthesis kit

The CFX96 andCFX384 system's solid-state optical technology (six filtered LEDs, eachwith a corresponding filtered photodiode) maximizes fluorescencedetection for specific dyes in specific channels, providing sensitivedetection for precise quantitation and target discrimination.

iScript™ cDNA Synthesis Kit

Bio-Rad's iScript cDNA Synthesis Kit | Biocompare …

The Aurum total RNA fatty and fibrous tissue kit produces high yields of exceptionally pure total RNA from samples that are difficult to disrupt. The kit is perfect for purifications involving fatty and fibrous tissues or samples such as fungi that are rich in RNases. It also works well with most animal and plant tissues, cultured cells, yeast, and bacteria. The Aurum total RNA fatty and fibrous tissue kit combines the best features of PureZOL™ RNA reagent, a potent phenol-based reagent that effectively lyses tissues and cells. Purification using PureZOL RNA reagent, with the speed of silica membrane technology in a spin-column format, yields high-quality, intact RNA in minimal time. The entire purification process, including DNase I digest, is completed in less than 60 minutes. The kit includes mini spin columns that can be processed using a choice of vacuum- or spin-mediated protocols. Isolated RNA is of the highest integrity and is suitable for the most discriminating applications.

iScript™  are formulated for efficient and sensitive reverse transcription over a wide linear dynamic range of input RNA for real-time qPCR.

iScript™ cDNA Synthesis Kit from Bio-Rad | SelectScience

The SensiFAST™ Probe One-Step Kits have been formulated for highly reproducible first-strand cDNA synthesis and subsequent real-time PCR in a single tube and uses a combination of the latest advances in buffer chemistry and enhancers, together with an antibody-mediated hot-start DNA polymerase system, to ensure fast, highly-specific and ultra-sensitive one-step RT-qPCR. This also gives SensiFAST Probe One-Step unbeatable efficiency in multiplexing. SensiFAST Probe One-Step Kits consists of a 2x SensiFAST Probe One-Step mix, separate reverse transcriptase and RiboSafe RNase Inhibitor.

iScript™ Select cDNA Synthesis Kit

5. After cDNA synthesis is complete, dilute samples to 100 µl each by adding 80 µl of RNase-free water. Samples are stable at -20°C or 4°C for a few weeks.