UltraRT cDNA Synthesis Kit
This kit is a cDNA first strand synthesis kit specially prepared for the first step experiment of two-step RT-PCR. The reverse transcriptase used in this kit is a novel and efficient reverse transcriptase that utilizes Escherichia coli engineering bacteria for recombination and expression. It removes RNase H activity and enhances its thermal stability. It can synthesize the first strand of cDNA using extremely low amounts of total RNA or mRNA, and the initial sample size can be as low as pg level. UltraRT reverse transcriptase has strong affinity for RNA and can read through RNA templates with high GC content and complex secondary structures, obtaining high yield cDNA.
This kit contains all the reagents required for reverse transcription from RNA templates to cDNA first strand, including UltraRT efficient reverse transcriptase, reaction buffer, primers, dNTP, etc., making it simple and convenient to use. This system has high compatibility with subsequent PCR and quantitative PCR tests, and is suitable for various DNA polymerase reactions.
Efficient reverse transcription: It has high affinity with RNA templates, with a reverse transcription efficiency of up to 90%, and can recognize pg level templates.
Free response to complex templates: Even templates with high GC content and complex secondary structures can achieve good results without high-temperature denaturation.
Component specifications
UltraRT (200U/ μ L) 100 μ L
five × UltraRT Buffer 500 μ L
Primer Mix 240 μ L
DNTP Mix, 2.5mM Each 500 μ L
RNase Free Water 1mL
Storage: -20 ℃, avoid repeated freeze-thaw.
During the operation process, RNase contamination should be avoided to prevent RNA degradation or cross contamination during experiments. It is recommended to perform RNA operations in specialized areas using specialized instruments and consumables. Operators should wear masks and disposable gloves and frequently change gloves.
Disposable plastic containers should be used as much as possible for experiments. If glass containers are used, they should be treated with a 0.1% DEPC (diethyl pyrocarbonate) aqueous solution at 37 ° C for 12 hours and sterilized under high pressure at 120 ° C for 30 minutes before use, or dry heat sterilized at 180 ° C for 60 minutes before use. The sterile water used in the experiment should be treated with 0.1% DEPC and subjected to high-pressure sterilization.
Before use, all reagents in this reagent kit should be gently mixed upside down to avoid foaming and used after brief centrifugation. The enzymes involved should be returned to -20 ℃ as soon as possible after use to avoid repeated freezing and thawing.
Attention: 1ng~5 μ Total RNA can be established by 20 μ L reaction system, if the total RNA content is greater than 5 μ g. Please expand the reaction system proportionally.
Reverse transcription steps:
Dissolve RNA templates, Primer Mix, dNTP Mix, UltraRT Buffer, UltraRT, and RNase Free Water and place them on ice for later use.
Prepare the reaction system according to the following table, with a total volume of 20 μ L.
Component dosage and final concentration
DNTP Mix, 2.5mM
