Lnc-RT Heroᵀᴹ I(With gDNase)(Super Premix for first-strand cDNA synthesis from lncRNA)
Specifications
25×20μl Rxns, 50×20μl Rxns
Lnc-RT HeroTM I (With gDNase) is a reverse transcription system specially developed for lncRNA to quickly remove genomic DNA contamination. 5×gDNase Mix can quickly remove the remaining genome in RNA at 42°C for 2 minutes, effectively avoiding the interference of genome on qPCR results.
5×L-RT HeroTM Mix contains Foregene LncRNA Reverse Transcriptase specially developed by Foregene, which is a new type of reverse transcriptase specifically for lncRNA and other long RNA complex templates, with stronger RNA affinity and higher reversal recording efficiency. The optimized system makes the reverse transcription rate faster and can easily transcribe RNA templates with high GC content and complex secondary structure. The first strand cDNA synthesis can be completed at 42°C in 15 minutes.
Kit components
| 5× gDNase Mix |
| 5× L-RT HeroTM Mix |
| RNase-Free ddH2O |
| Instructions |
Features&advantages
■ Efficient ability to remove gDNA, which can remove gDNA in the template within 2 minutes.
■ It can efficiently reverse transcription of lncRNA. When the reverse transcription product is subjected to qPCR, the Ct value is lower than that of conventional reverse transcription system.
■ Efficient reverse transcription system, it only takes 15 minutes to complete the synthesis of the first strand cDNA.
■ Complex templates: templates with high GC content and complex secondary structure can also be reversed with high efficiency.
■ High-sensitivity reverse transcription system, pg-level templates can also get high-quality cDNA.
■ The reverse transcription system has high thermal stability, the optimal reaction temperature is 42℃, and it still has good reverse transcription performance at 50℃.
Kit application
lncRNA relative quantitative analysis.
lncRNA absolute quantitative analysis.
It can quickly and accurately analyze trace RNA such as RNA virus.
Reverse transcription of RNA templates with high GC content or complex secondary structure.
Work flow
Storage and Shelf life
The kit should be stored at -20°C. Store the product in a constant temperature refrigerator at -20°C immediately after receipt. If the storage conditions are appropriate, the product will not degrade any performance during the 1-year validity period.
Problem Analysis Guide
The following is an analysis of the problems that may be encountered in the use of Lnc-RT HeroTM I (With gDNase) series kits, hoping to be helpful to your experiments. In addition, we have dedicated technical support to help you with other experimental or technical problems beyond the operating instructions and questions. If you have any needs, please contact us: 028-83361257 or E-mail: Tech@foregene.com.
Non-specific amplification
1. Unreasonable primer design
Recommendation: Design primers according to primer design principles.
2. Genome residues.
Suggestion: Make sure that the first step of gDNA removal reaction temperature is 42°C, and the reaction time can also be extended to 5min.
No amplification signal by RT-qPCR
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 the 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 when handling to prevent DNA samples from being sucked into the pipette or spilled out of the centrifuge tube.
2. Contamination occurred during the preparation of the PCR reaction system.
Recommendation: Take necessary precautions when handling, such as: wearing latex gloves, using filter tips. Use Real Time PCR Mix in a contamination-proof system.
3. The primers are degraded.
Suggestion: Use SDS-PAGE electrophoresis to detect whether the primers are degraded, and replace with new primers for fluorescence detection experiments.
Instruction Manual:
