Lab Reagent Reverse Transcription Kits RT EASY II For Generating First Strand CDNA materials and equipment lab reagent

Fast and highly sensitive lab reagent reverse transcription kits RT EASY II for generating first-strand cDNA

Kit application

Directly used for Real Time PCR quantitative analysis of gene expression.

Can quickly and accurately analyze trace amounts of RNA such as RNA viruses.

Reverse transcription of RNA templates with high GC content or complex secondary structure.

Kit Component Information

2×RT OR-Easy^TM Mix: Foregene Reverse Transcriptase, RNase Inhibitor, dNTPs, Stabilizer, Enhancer, Optimizer and Reverse Transcription Primer (Random Primer, Oligo(dT)₁₈Primer) with optimized ratio.

RT Mix tolerance

The 2×RT OR-Easy^TM Mix showed a very high tolerance to alcohol and guanidine salts after testing analysis. The RNA purified in the laboratory often has residues of alcohol and guanidine salts, which have a strong inhibitory effect on reverse transcription, resulting in unsatisfactory reverse transcription effect or low reverse transcription efficiency. The tolerance of Foregene RT Easy series products to alcohol and guanidine salts makes reverse transcription easier and more efficient.

Analysis of Alcohol Tolerance

Analysis of guanidine salt tolerance

Preparation before operation

It is strongly recommended that users read the instructions carefully before using this kit. RT Easy series kits are easy to operate, convenient and fast, and the instructions provide the correct use of the entire kit. Please prepare necessary experimental materials and equipment before use.

Template RNA amount

Template RNA should preferably be extracted from fresh samples or stored at -80°C (repeated freezing and thawing of RNA should be avoided).

RT EasyTM II: (0.1pg-0.5μg total RNA or 0.01pg-0.05μg mRNA)/10μl system.

Nonspecific amplification

1. Unreasonable primer design.

Recommendation: Design primers according to primer design principles.

2. Genomic residues.

Recommendations: Use a reverse transcription kit such as (Cat. No. RH-01031) that contains genome ablation or design trans-intron primers.

RT-QPCR no amplification signal

1. RNA is degraded.

Recommendation: The material for RNA extraction should be as fresh as possible, and high-quality and high-purity RNA should be used.

2. RNA contains inhibitors.

Recommendation: Reverse transcription inhibitors generally include SDS, guanidine salts, EDTA, etc. It is recommended to wash the RNA precipitate with 70% ethanol to remove inhibitors.

3. Primer design issues.

Recommendation: According to the primer design principle, redesign the primers for inspection.

The target band appears in the blank control

1. Contamination of operating tools or reagents.

Recommendation: All reagents or equipment for the experiment should be autoclaved. Be careful and gentle to prevent DNA samples from being drawn into the pipette or spilled out of the centrifuge tube.

2. Contamination occurred during PCR reaction system preparation.

Recommendation: Take necessary precautions when handling, eg: Wear latex gloves, Use filter tips. Use real time PCR Mix in a contamination-proof system.

3. The primers are degraded.

Recommendation: Use SDS-PAGE electrophoresis to detect whether the primers are degraded, and replace with new primers for fluorescence detection experiments.