Product Number: PCK34

Shipping and Storage

Stored at -20℃, valid for 12 months.

Component

Description

This kit is specifically designed for one-step RT-PCR experiments, and can be used to conveniently and quickly complete reverse transcription and PCR amplification reactions in the same reaction tube. During the reaction process, there is no need to open the tube cap to add reagents, which avoids contamination while improving detection sensitivity and experimental efficiency. This kit includes the OneStep Enzyme Mix optimized for optimal ratio (including mutated TRUEscript M-MuLV H Minus reverse transcriptase, hot start HotMaster Taq DNA polymerase, and RNasin inhibitor mix), as well as a unique 2 × RT-PCR Mix reaction system suitable for reverse transcription and PCR amplification. The RNase H activity of this enzyme M-MuLV (RNase H ˉ) is deficient. Compared with M-MuLV, it has stronger extensibility and stability, and can be used for longer cDNA synthesis and the construction of high proportion full-length cDNA libraries. At the same time, the enzyme enhances heat resistance and can reverse transcribe at 42-50 ° C, improving the efficiency of complex secondary structures and GC rich template reverse transcription.

Application

Suitable for high copy and low copy gene testing; RNA templates with high GC content or complex secondary structures.

Product Number: RT07

Shipping and Storage

Store at -30 ~ -15 ℃ and transport on dry ice.

Components

Description

Super FiV M-MLV Reverse Transcriptase (RNase H-) is a reverse transcriptase that recombines and expresses mutated M-MLV genes in E. coli engineering bacteria. This enzyme can catalyze complementary DNA polymerization reactions using RNA or DNA: RNA hybrid chains as templates. The mutated Super FiV M-MLV Reverse Transcriptase (RNase H-) RNase H activity is missing, reducing RNA degradation in reverse transcription reactions and making it easier to obtain full-length cDNA. Super FiV M-MLV Reverse Transcriptase (RNase H-) exhibits excellent reverse transcription activity at 55 ℃ (the enzyme can be used for reverse transcription reaction at up to 60 ℃). For complex RNA structures, increasing the reverse transcription reaction temperature can significantly improve the yield of cDNA. In addition, the mutated Super FiV M-MLV Reverse Transcriptase (RNase H-) has better stability and can synthesize up to 15kb of cDNA. Suitable for the synthesis of first stranded cDNA, RT PCR, RT qPCR, and construction of full-length cDNA libraries.

Unit definition

Using Poly (A) as a template and oligo (dT) as a primer, the enzyme required to catalyze the addition of 1 nmol of dTTP within 10 minutes at 37 ℃ is defined as one active unit (U).

Quality control

200 U of this enzyme reacted with 1µg of 16S, 23S rRNA at 37 ℃ for 1 hour, and the electrophoresis band of the RNA remained unchanged.

Product Number: PCK5802

Shipping and Storage

Low temperature transportation, stored at -20℃, with a shelf life of one year.

Component

Description

This kit provides all the reagents required for synthesizing the first strand of cDNA. The principle is to efficiently synthesize full-length cDNA copies of mRNA using mutated Moroni murine leukemia virus reverse transcriptase (M-MuLV RT).

Features

1. The use of mutated reverse transcriptase results in low endogenous RNase H activity.
2. Can synthesize cDNA with a length of 9kb or more.
3. Oligo dT and random primers can be used, which are provided in this kit.
4. Total RNA, poly (A)+RNA, or special RNA can all be used as templates.
5. Can be used for the synthesis of the second chain RT-PCR, Probe labeling, etc.

Product Number: RT10

Shipping and Storage

Storage at -25~-15°C and transportation≤ 0℃. Avoid repeated freezing and thawing, and valid for 18 months.

Component

Description

X Reverse Transcriptase is  an  aptamer-modified  and RNA-template-dependent DNA polymerase that inhibits its reverse transcription activity below 40°C. X Reverse Transcriptase lacks 3′ → 5′ exonuclease activity and has RNase H activity. The enzyme can use RNA as a template to synthesize a complementary DNA strand, which can be applied to first-strand cDNA synthesis. Because of its high activity between 50-65°C, it is particularly suitable for RT-LAMP (Loop-Mediated Isothermal Amplification). Since the polymerase activity of X Reverse Transcriptase is inhibited below 40°C, the non-specific amplification caused by mismatching of primers during the preparation of the reaction system can be greatly reduced, solving the problems of primer-dimer formation before the reaction, improving the consistency and specificity of amplification.  X Reverse Transcriptase  supports high-throughput applications and operation at room temperature. X Reverse Transcriptase (Glycerol free) can be used to prepare lyophilized  preparations,  lyophilizable RT-LAMP reagents, etc.

Application

1. cDNA synthesis;
2. Combined with Bst DNA;
3. RT-LAMP isothermal amplification can be performed to detect target RNA.

Unit definition

One unit incorporates 1nmol of dTTP into acid-precipitable material in 20 minutes at 50°C using poly(A)•oligo(dT) as template-primer.

Shipping and Storage

2-8℃.

Component

Note

1. Usually, a primer concentration of 0.2μM can yield good results, and a reference range of 0.1-1.0μM can be used for setting. The concentration of the probe used is related to the fluorescent quantitative PCR instrument, probe type, and fluorescent labeling substance used. Please refer to the instrument manual or the specific usage requirements of each fluorescent probe for concentration adjustment during actual use.
2. Usually, the amount of RNA template is based on 10pg-100ng as a reference. Due to the different copy numbers of the target gene contained in templates of different species, gradient dilution of the template can be performed to determine the optimal template usage.

Product Number: PCK53

Storage condition

Store at -20°C.

Component

Description

The MicroRNA cDNA Kit for First Chain cDNA Synthesis is a specialized kit based on the principle of the stem loop method. The 5 × THERMO Reaction Mix is a tubular reverse transcription premix, containing all the reagents required for reverse transcription (THERMScript H-RTase, RNase inhibitor, dNTP Mixture, Buffer). Simply add template RNA and stem loop primers for the reaction, making cDNA synthesis more convenient and efficient. This kit uses a special gDNA Remover with DNA degradation activity, which does not require opening the lid halfway to add. With just one step, genome clearance and reverse transcription reactions can be completed simultaneously, greatly simplifying the operation steps and avoiding the risks of sample contamination and RNA degradation caused by complex sample addition processes. This product is based on THERMOscript H ˉ RTase, which has extremely high thermal stability. Coupled with an optimized buffer system, it maximizes the synthesis of various microRNA specific reverse transcription products. The cDNA product has good compatibility and can be paired with any commercial company’s fluorescence quantitative mix for downstream detection experiments.

Product Number: PCK52

Storage condition

Store at -20°C .

Component

Description

The enhanced microRNA reverse transcription/fluorescence quantitative detection kit contains all the reagents for microRNA detection. This product adopts the Poly (A) tail addition method, using microRNA as a template, and using a specially optimized pre mixed microRNA RT Enzyme mix (including Poly (A) tail addition enzyme and reverse transcription enzyme) to efficiently complete cDNA synthesis through a one-step method of Poly (A) tail addition and reverse transcription; MicroRNA detection was performed using a 2 x microRNA qPCR mix. Suitable for samples containing microRNAs such as Total RNA or Small RNA.

Features

1. The optimal ratio of Poly (A) tail enzyme and reverse transcriptase, as well as the optimized reaction buffer, ensure the reverse transcription efficiency of microRNA.
2. PolyA tail addition and reverse transcription cDNA synthesis are completed in one step in the same tube.
3. 2×microRNA qPCR Mix has high amplification efficiency, strong specificity, and sensitivity.
4. Equipped with ROX Reference Dye, it can be used for various models that require high and low ROX reference dyes.

Product Number: PCK51

Storage condition

Store at -20°C in the dark for at least 12 months, thoroughly dissolve and mix before use. 2×microRNA qPCR Mix (with Sybr Green) can be stored at 4°C for short-term use to avoid repeated freeze-thaw cycles. Reverse primer (10μM) should be stored at -20°C after each use.

Component

Description

         The enhanced microRNA fluorescence quantitative PCR detection kit adopts SYBR The principle of Green I chimeric fluorescence method for microRNA fluorescence quantitative detection. This kit contains all the reagents for microRNA fluorescence quantitative detection, including 2×microRNA qPCR Mix and Reverse Primer. 2×microRNA qPCR Mix (including Sybr Green) is a new generation of pre mixed fluorescent quantitative PCR detection reagent specially developed for microRNA quantitative detection. The DNA polymerase used in it is an antibody modified hot start form, combined with a special buffer system, making the reaction more specific, sensitive, and able to accurately quantify over a wider range.

Note: This kit must be used in conjunction with the Enhanced microRNA Real-Time PCR Assay kit(PCK50).

Product Number: PCK50

Storage condition

-20°C

Component

Description

         Enhanced microRNA 1st Strand cDNA synthesis kit This kit uses the Poly (A) tail addition principle. Firstly, a Poly (A) tail is added to the 3 ‘end of microRNA, and then an Anchored oligo (dT) – universal tag universal reverse transcription primer is used for reverse transcription reaction to generate the corresponding cDNA first chain of microRNA. This kit uses a specially optimized pre mixed microRNA RT Enzyme Mix to combine Poly (A) tailing and reverse transcription into one step, simplifying the operation steps and improving the efficiency of Poly (A) tailing and reverse transcription. The kit has the ability to effectively prepare microRNA corresponding cDNA first strands from 20pg-2μg Total RNA. A single synthesized cDNA can detect multiple microRNAs, saving samples and costs.

Note: This kit must be used in conjunction with the Enhanced microRNA Real-Time PCR Assay kit(PCK51).

Product Number: RT5060

Shipping and Storage

Store at -20℃.

Components

Description

ThermoPlus M-MLV Reverse Transcriptase(RNase H-),encoded by Moloney Murine Leukemia Virus (M-MLV RT) is an RNA-dependent DNA polymerase that synthesizes the complementary cDNA first strand from a single-stranded RNA template to which a primer has been hybridized. MMLV RT will also extend primers hybridized to single-stranded DNA. Second strand cDNA synthesis can be achieved from some mRNA templates without an additional DNA polymerase. thermol plus MMLV is a mixture of several temperature-resistant reverse transcriptase enzymes with good temperature adaptability. The optimal temperature range is 50-60 ℃, adapting to a variety of different reaction conditions. It has excellent reverse transcription performance.

Source

Recombination of E.coli containing Moloney murine leukemia virus reverse transcriptase gene from clone of Moloney murine.

Concentration

200U/μl

Features

1. Lack RNase H activity: Weak RNaseH activity High cDNA yield, can get more full length cDNA.
2. Thermal stable: the optimum reaction temperature is 50℃,the highest is 60℃. Can overcome the template RNA secondary structure ,and finish the reverse transcriptase experiment smoothly.
3. Wide temperature range: can reverse transcript from 37-60C,with more than 80% of the highest activity at 42℃-55℃ customer can choose the reaction temperature freely.
4. Strong amplification activity: Gene mutation enhanced the binding capacity of the enzyme and RNA.Increased the amplification speed, can obtain the quality cDNA, suitable for cDNA library construction.

Application

The first-strand cDNA synthesis; RT-PCR.

Unit definition

One unit of MMLV RT catalyzes the incorporation of 1 nmol of dTTP into acidinsoluble material in 10 minutes at 37℃ using oligo(dT)12-18-primed poly(A)n as a template.

Storage buffer

20 mM Tris-HCl (pH7.5),200 mM NaCl, 0.25 mM EDTA,0.01% NP-40(v/v),2.5 mM DTT,50% glycerol (v/v).

5×RT Buffer:250mM Tris-HCl (pH 8.3), 15mM MgCl2,375 mM KCl,50mM DTT.