lncRNA Reagents
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Features
● High efficiency: Stable and efficient King reverse transcriptase with RT efficiency over 95%.
● Sensitivity: The sensitivity-enhancing components are added to the buffer solution, ensuring the high sensitivity with lower limit of detection reaching 0.1 ng total RNA.
● Stress resistance: Suitable for complex templates, perfect resistance to foreign interference.
● Fast reaction: gDNA-free cDNA can be synthesized within 21 min.
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Description
The lnRcute lncRNA First-Strand cDNA Kit is a product specially developed for reverse transcription of long-chain non-coding RNA (lncRNA). Compared with mRNA, lncRNA has the characteristics of low abundance, large difference in GC content and more complex secondary structure, and traditional reverse transcription reagents are difficult to achieve ideal effects on lncRNA. This kit contains gDNase for efficiently removing genomic DNA, which can effectively avoid interference of residual genomic DNA on subsequent detection results. The reverse transcriptase used in lnR RT Enzyme Mix in this kit is FastKing RT Enzyme. This enzyme is a new molecular modified reverse transcriptase, with a hydrophobic motif added, and has stronger RNA affinity and thermal stability, which makes this kit outstanding in aspects of stress resistance and high reverse transcription efficiency for templates with GC content, complex secondary structure or low abundance. This kit is especially suitable for reverse transcription reaction of lncRNA with relatively low expression level and relatively complex secondary structure.
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Kit Contents
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Applications
The reverse transcribed cDNA can be used for fluorescent quantitative PCR of lncRNA, and for routine PCR of mRNA, fluorescent quantitative PCR, cDNA library construction and other experiments. -
Storage Condition
Store at -30~-15°C for one year. -
Important Notes
● The following protocol is applicable to the total RNA with the template amount of 10 ng-2 μg. If the total RNA amount is higher than 2 μg, please increase the reaction system volume in proportion.
● Please operate on ice to prevent RNA degradation.
● For RNA template with complex secondary structure, a denaturing step is recommended, that is, before starting, incubate the template RNA at 65°C for 5 minutes, then transfer to ice quickly to perform the next step.
● According to different experimental needs, the Oligo-dT Primer or Gene Specific Primer can also be applied. The amount of primers is as follows: Oligo-dT Primer 50 pmol/20 μl reaction system, Gene Specific Primer 5 pmol/20 μl reaction system.
● The non-specific amplification effect during the PCR reaction can be decreased by improving the reverse transcription reaction temperature to 50°C.
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