| Product Description | This product is a mixture of fast reverse transcription reagents. The 5 x EasyQuick RT MasterMix contains all the reagents required for reverse transcription from RNA templates to cDNA first strand, including EasyQuick RT Reversase, RNase Inhibitor, Random 6 mers, Oligo dT Primer, dNTP, EQ-RT Buffer, etc. The reverse transcription efficiency of this product is high, and it can perform a good reverse transcription reaction on a small amount of RNA templates. The fluorescence quantitative template cDNA first strand synthesis can be completed in 15 minutes. This reagent kit is very convenient and fast to operate, and only RNA templates and water need to be added for reverse transcription reaction, making it particularly suitable for high-throughput detection. | E665905 | Component | 200 T | Storage | | E665905A | 5×EasyQuick RT MasterMix | 400 μL | -20℃. Avoid freeze/thaw cycle. | | E665905B | RNase-Free Water | 2×1 mL | -20℃. Avoid freeze/thaw cycle. |
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Product features
1. Convenience: The ready to use reverse transcription Mix only requires the addition of RNA templates and water to initiate the reaction.
2. Fast: Complete cDNA first strand synthesis in 15 minutes.
3. High reverse transcription efficiency: The reverse transcription efficiency is above 90%.
4. High sensitivity: PG level templates can also obtain high-quality cDNA.
5. Read through complex templates: templates with high GC content and complex secondary structures. Matters needing attention
1. During the operation, RNase contamination should be avoided to prevent RNA degradation or cross contamination during experiments. It is recommended that operators wear masks and disposable gloves, frequently change gloves, and use specialized instruments and consumables.
2. The reverse transcription system is prepared on ice for operation to prevent RNA degradation. The MasterMix of the reagent kit should be stored at -20 ℃ as soon as possible after use, and repeated freeze-thaw should be avoided as much as possible.
3.10 μ The reaction system can be used up to 1 μ G Total RNA, if the amount of template RNA is greater than 1 μ g. Please expand the reaction system proportionally.
4. For RNA templates with complex secondary structures, it is recommended to incubate the template RNA at 65 ℃ for 5 minutes on ice before proceeding with the next step, followed by brief centrifugation. Operation steps
1. Thaw the template RNA on ice; After thawing the components of the reagent kit at room temperature, immediately place them on ice. Before use, vortex shake and mix each solution, and centrifuge briefly before use.
2. Prepare the reaction system according to the following table (please prepare the reaction solution on ice), vortex shake and mix well, briefly centrifuge, and collect the solution on the tube wall to the bottom of the tube.
| Reagent |
10 μl Reaction system |
Final concentration |
| RNA Template |
X μl |
1 pg～0.5 µg ¹⁾ |
| 5×EasyQuick RT MasterMix ²⁾ |
2 μl |
1× |
| RNase-Free Water |
up to 10 μl |
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Attention:
1) If the total RNA content is greater than 1 µ g, please expand the reaction system proportionally.
2) 5 x EasyQuick RT MasterMix contains Oligo (dT), Random Prime, RNase Inhibitor, dNTP Mixture, EQ-RT Buffer, etc.
3.
Incubate at 37 ℃ for 15 minutes.
4.
Incubate at 85 ℃ for 5 seconds to inactivate reverse transcriptase.
5. After a brief centrifugation, place it on ice for subsequent experiments. If it needs to be stored for a long time, please place it at -20 ℃.
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