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LiScript™ First-Strand cDNA Synthesis Kit (+gDNA wiper) is a kit for reverse transcription after removal of genomic DNA. The kit removes genomic DNA in 2 minutes at 42°C. Meanwhile, because the reverse transcription reagents contain components that inhibit gDNA Eraser, the sample processed by gDNA Eraser can be directly used for reverse transcription reaction to synthesize cDNA.
The kit contains a novel high-performance LiScript™ reverse transcriptase. The novel mutation site greatly enhances the transcriptional activity of the enzyme. The efficiency and yield of cDNA first-strand synthesis are higher, and the first strand of cDNA can be synthesized using pg total RNA or mRNA. If the cDNA is used for downstream qPCR, the reverse transcription reaction can be completed at 42°C for 15 minutes. This kit is suitable for the synthesis of first-strand cDNA and subsequent RT-PCR, RT-qPCR, and construction of full-length cDNA libraries.
Key Features
• Rapid genomic DNA deletion: With the gDNA Eraser, it takes only 2 minutes to remove genomic DNA.
• Rapid reverse transcription: It takes only 15 minutes to obtain the first strand of cDNA.
• High sensitivity: pg of total RNA or mRNA can be used as template.
• High efficiency of reverse transcription efficiency: the novel mutation site enhancers the activity of the enzyme, to increasethe yield of cDNA.
Components
1. gDNA Eraser: 50 μl
2. 10× gDNA Erase Buffer: 120 μl
3. LiScript™ Reverse Transcriptase: 100 μl, 200 U/μl
4. 5× ScriptRT Buffer: 500 μl
5. Primer Mix: 120 μl
6. RNase-Free Water: 2×1 ml
Storage
Store at -20°C.
Case Study
cDNA fragments as long as 20 kb was successfully obtained from RNA templates using LiScript™ 1st Strand cDNA Synthesis Kit (Left Figure). Reverse transcription was performed using 1 μg total RNA of HeLa cells or total RNA (for Nebulin) of skeletal muscle cells as templates and Oligo(dT)23VN as primers. PCR amplification was performed using 1 μl of cDNA (as template) and DNA Polymerase.
LiScript™ has higher reverse transcription efficiency for complex RNA templates (Right Figure). Using 1 μg of total RNA of Hela cells as template, reverse transcription was performed using LiScript™ and a similar product of T brand, respectively. The reaction time is 5 min. Using 1 μl of cDNA as template, the high-GC regions of FIB (6100 bp) and Pole (7117bp) were amplified using LiQuant™ Green qPCR Master Mix, respectively.
LiScript™ has a wide and sensitive linear reverse transcription range for qPCR. Reverse transcription was perform using 1 μg-1 pg total RNA of Hela cells as template and random hexamers/Oligo(dT)23VN as mixed primers. qPCR amplification of amplify ACTB gene and BR3 gene was performed using 1 μl of cDNA (as template) and LiQuant™ Green qPCR Master Mix, respectively.
