• One-tube RT Kit
  • One-tube RT Kit

One-tube RT Kit

1. The synthesis of cDNA is more convenient, and users only need to prepare templates and water for the reaction. 2. It is fast and the operation steps have been simplified to the maximum extent, which can reduce pollution and experimental errors
$1.70
  • One-tube RT Kit

SPECIFICATION


                                                One-tube RT Kit
This product is a one tube reverse transcription premix mix that contains all the reagents required for reverse transcription first chain synthesis (MMLV, Random primers, Oligo dT Primer, dNTP Mixture, Reaction Buffer, etc.). Users only need to add template RNA and RNase Free water to perform the reaction.
Product features:
1. The synthesis of cDNA is more convenient, and users only need to prepare templates and water for the reaction.
2. It is fast and the operation steps have been simplified to the maximum extent, which can reduce pollution and experimental errors.
3. High efficiency, specially optimized oligo dT and N6 random primer ratio, high reverse transcription efficiency.
4. Fast speed, it only takes 42 ℃ and 20 minutes to complete the synthesis of the first strand of cDNA.
5. It can be used for RNA templates with high GC content and complex secondary structures, and can synthesize cDNA fragments with a length of up to 12kb.
6. Good compatibility, the synthesized cDNA can be directly used for fluorescence quantitative PCR reaction, with high sensitivity and good stability.
Kit composition:
Component specifications
two × RT MasterMix 0.1ml
RNase Free water 1ml
1 instruction manual
Storage conditions: Low temperature transportation, -20 ℃ storage, valid for one year.
Usage:
1. After thawing, flip the dough upside down and mix all components evenly, using 20 μ Taking L system as an example, add (2) in the following table × RT Master Mix is very viscous, please centrifuge briefly before use and avoid loss of adhesion to the outer wall of the suction head):
two × RT MasterMix 10 μ L
Total RNA/mRNA template (self prepared) 50ng-2 μ G/5-200ng
Replenish RNase Free water to 20 μ L
Note: For RNA templates with complex secondary structures, it is recommended to use the denaturation step, which involves incubating the template RNA at 65 ℃ for 5 minutes before quickly transferring it to ice before proceeding to the next step.
2. Gently mix and incubate at 42 ℃ for 20 minutes for reverse transcription reaction.
Note: For templates with complex secondary structures or high GC content, the temperature can be raised to 50 ℃ to enhance reverse transcription efficiency.
Heat at 3.85 ℃ for 5 seconds to inactivate reverse transcriptase and store on ice for later use (please store at -20 ℃ for longer periods).
Precautions:
During the experiment, please pay attention to avoiding RNase contamination and preventing RNA degradation or cross contamination during the experiment.
2. Before use, please completely dissolve and mix thoroughly to prevent uneven salt ion concentration from affecting the experimental results.
3. After use, it should be returned to -20 ℃ as soon as possible to avoid repeated freezing and thawing.
4. The integrity of RNA templates plays a decisive role in the efficiency of cDNA synthesis, so please choose a reliable RNA extraction/purification method.
If the amplified fragment is long or the RNA structure is complex, in order to enhance the transcription effect, the RNA can be separately heated at 65-70 ℃ for 5-10 minutes before being added to the system.
6. Operators should wear masks and disposable gloves, and
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