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0In the majority of instances, genomic DNA (gDNA) may be present in extracted RNA. If the mixed gDNA is not removed, the reverse-transcribed cDNA product will likewise be contaminated with gDNA. If qPCR primers are not designed to span introns, cDNA and gDNA may be concurrently amplified in the qPCR reaction, leading to biased or even meaningless cDNA quantitative data.
gDNA removal is typically conducted on extracted RNA samples, allowing for more precise experimental results. gDNA removal is often performed before to reverse transcription. After gDNA removal has been completed under specific conditions, reverse transcription-related reagents will be added. This technique is more difficult, and frequent cap opening and sample addition will increase to some degree. Possibility of sample contamination and degeneration.
Yeasen Biotechnology's Hifair™ V one-step RT-gDNA digestion SuperMix for qPCR (Cat# 11142) can execute reverse transcription and gDNA removal in the same tube, making the operation simpler and the experiment more secure!
Features
Simple operation: one-step reverse transcription and genome removal.
Efficient removal: excellent removal of 200 ng genome.
Excellent synthesis: GC-rich and low-GC genes reverse transcribe well.
High sensitivity: 1pg sensitivity, high sample detection rate.
High stability: the kit can be stored at 4°C for 14 days and the cDNA for 14 days at 37°C with 50 freeze-thaw cycles.
Wide application: human, mouse, Arabidopsis, E. coli, etc.
Product Showcase
1. Excellent gDNA removal ability
Figure 2. Add 200 ng of 293T gDNA to the amplification system, and use Yeasen, A* brand, B* brand, and control components for gDNA removal. qPCR was used to quantify the cDNA acquired during reverse transcription. The results indicate that Yeasen products are highly effective at removing gDNA.
2. Genes with 30%-70% GC reverse transcribe well
Figure 3. Using the total RNA of 293T cells as a template, reverse transcription was done using Yeasen, A* brand, and the resulting cDNA was analyzed quantitatively by qPCR. Results indicate that the reverse transcription efficiency of Yeasen products is superior to that of imported brands.
3. High detection rate, up to 1pg
Figure 4. Using 1 pg-1 g (7 gradients) of 293T cell total RNA as a template, reverse transcription was done by Yeasen, A* brand, and B* brand products, and the resulting cDNA was quantitatively evaluated using qPCR. Results indicated that the detection rate of 1 pg-1 ug for Yeasen goods was superior to that of competing brands.
4. The product is stable after 14 days at 4°C
Figure 5. Using the total RNA of 293T cells as a template, reverse transcription was done using kits maintained at 4°C for 14 days and at -20°C, and the resultant cDNA was evaluated quantitatively by qPCR. The results indicate that the Yeasen kit is still stable after 14 days of storage at 4°C. Note: CT = CT (stored at 4°C for 14 days) - CT (stored at -20°C). CT is within 0.5, and the difference between the two is deemed insignificant.
5. The cDNA product has good stability
Figure 6. Using the total RNA of 293T cells as a template and Yeasen Cat#11142 for reverse transcription, cDNA was produced and stored at 37 °C for 3 days, 7 days, and 14 days before being tested 10 times, 30 times, and 50 times. As a control, cDNA was kept at -20°C. Quantitative qPCR analysis was conducted on cDNA following various treatments. The results indicated that the CT obtained by quantifying cDNA under different conditions was within the range of 0.1, and the product exhibited outstanding stability.
Note: CT = CT (cDNA processed under different conditions) - CT (cDNA stored at -20°C). CT is within 0.5, and the difference is regarded as trivial.
6. Wide range of species application
Figure 7. Using 1 g/10 ng of total RNA from Arabidopsis thaliana, mouse, and Escherichia coli as a template, reverse transcription was done with Yeasen, A* brand, and B* brand products, and qPCR was used to assess the generated cDNA. The results indicate that the amplification CT value of Yeasen goods in various templates is less than or similar to that of other brand products and has high efficiency of reverse transcription.
Feedback
👉Peking University Shenzhen Graduate School
RNA sample: from THP-1 cells, the RNA concentration is 100 ng/μL
Customer evaluation: One-step method is very easy to use
👉Nanjing Agricultural College
RNA sample: from soybean total RNA, the RNA concentration is 300 ng/μL
Customer evaluation: This product has drastically reduced the number of experimental stages and duration, while significantly enhancing the experimental efficiency.
Ordering Information
|
Product Name |
Product Code |
Specification |
|
Hifair™ V one-step RT-gDNA digestion SuperMix for qPCR |
11142ES10 |
10 T |
|
11142ES60 |
100 T |
Related Products
|
Product Positioning |
Product Name |
Product Code |
|
qPCR Mix |
11184ES |
|
|
Reverse Transcriptase |
Hifair™ III Reverse Transcriptase (200U/μL) |
11111ES |
|
11300ES |
