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: 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.

Product Number:RT05F

Shipping and Storage

Stored at -30~-15℃, Dry ice transportation to avoid repeated freezing and thawing.

Components

Description

HiFi II M-MLV Reverse Transcriptase (RNase H-) (Glycerol-free) is the glycerol free version of HiFi II M-MLV Reverse Transcriptase (RNase H-),it is a reverse transcriptase that recombines and expresses the mutated M-MLV gene using Escherichia coli engineering bacteria,this enzyme can catalyze complementary DNA polymerization reactions using RNA or DNA: RNA hybrid chains as templates.The mutated HiFi II M-MLV (H -) reverse transcriptase RNase H activity is missing, reducing RNA degradation in reverse transcription reactions and making it easier to obtain full-length cDNA.HiFi II M-MLV (H -) reverse transcriptase can synthesize the first strand of cDNA at 55℃, providing higher specificity and stability. It can synthesize up to 12kb of cDNA with high cDNA yield.Suitable for the synthesis of first strand cDNA, RT-PCR, RT-qPCR, construction of full-length cDNA libraries, and application of freeze-dried RT-PCR products.This product does not contain freeze-dried shaping ingredients, and can be customized and added according to needs when applied to freeze-dried products.

Unit definition

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

Quality Control

The electrophoresis bands of the RNA did not change when 200U of this enzyme reacted with 1µg of 16S, 23S rRNA at 37 ℃ for 1 hour.

Notes

1. 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.
2. 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℃ for 12 hours and sterilized under high pressure at 120℃ for 30 minutes before use, or dry heat sterilized at 180℃ 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.
3. 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.
4. If the initial amount of RNA is less than 50ng, it is recommended to add RNA enzyme inhibitors (RNase inhibitor, Murine).

Product Number: RT05

Shipping and Storage :

-20°C

Components

Description

HiFi II M-MLV(H-) is a reverse transcription enzyme in which the mutant M-MLV gene is recombined and expressed by Escherichia coli engineering bacteria. The enzyme can catalyze the polymerization of complementary DNA using RNA or DNA: RNA hybrid chain as template. The mutation of HiFi II M-MLV reverse transcriptase RNase H activity is lost, reducing the degradation of RNA in reverse transcription and facilitating the acquisition of full-length cDNA. HiFi II M-MLV reverse transcriptase can synthesize the first strand cDNA at 55°C, providing higher specificity and stability, and can synthesize up to 12 KB cDNA with high cDNA yield. It is suitable for synthesis of first strand cDNA, RT-PCR, RT-QPCR and construction of full-length cDNA library.

Activity Definition

Using Poly (A) as template and Oligo (dT) as primer, the amount of enzyme required for catalytic incorporation of 1 nmol dTTP within 10 minutes was defined as an activity unit (U) at 37°C.

Quality Control

The electrophoretic bands of RNA did not change after the reaction of 200 U of this enzyme with 1 μg of 16 S and 23 S rRNA at 37°C for 1 hour.

Product Number: RT04

Storage condition

-20°C

 Component

Description

MMLV Reverse Transcriptase, encoded by Moloney Murine Leukemia Virus (MMLV 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. The difference between this to the general MMLV RT is that the capacity to endure the heat is enhanced. It can remain the 100% activity at 50°C, it can also keep more than 80% activity even at 60°C.

Features

1. Lack RNase H activity: Weak RNase H activity High cDNA yield, can get more full-length cDNA.
2. Thermal stable: the optimum reaction temperature is 50°C, the highest is 60°C. Can overcome the template RNA secondary structure, and finish the reverse transcriptase experiment smoothly.
3. Wide temperature range: can reverse transcript from 37-60°C, with more than 80% of the highest activity at 42°C-60°C. 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.

Source

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

Unit definition

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

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×MMLV Reaction Buffer:

         [5×MMLV Reaction Buffer] 250mM Tris-HCl (pH 8.3), 15mM MgCl2, 375 mM KCl, 50mM DTT.

Product Number: RT03

Storage condition

-20°C

Component

Description

MMLV Reverse Transcriptase, encoded by Moloney Murine Leukemia Virus (MMLV 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. The difference between this to the general MMLV RT is that the capacity to endure the heat is enhanced. It can remain the 100% activity at 50°C, it can also keep more than 80% activity even at 55°C.

Features

• Lack RNase H activity: Weak RNase H activity High cDNA yield, can get more full-length cDNA.
• Thermal stable: the optimum reaction temperature is 50°C, the highest is 55°C. Can overcome the template RNA secondary structure, and finish the reverse transcriptase experiment smoothly.
• Wide temperature range: can reverse transcript from 37-55°C, with more than 80% of the highest activity at 42°C-55°C. customer can choose the reaction temperature freely.
• 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.

Source

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

Application

The first-strand cDNA synthesis; RT-PCR.

Unit definition

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

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×Reaction Buffer:

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

Product Number: RT01

Storage condition

-20°C

Component

Description

         MMLV Reverse Transcriptase isolated from E. coli carrying rat leukemia virus pol gene consists of a single peptide, molecule weight 71kd. MMLV Reverse Transcriptase, RNase H Minus is an RNA-dependent DNA polymerase with no detectable RNase H activity. A point mutation in the RNase H domain increases the thermostability of the enzyme and support greater cDNA yield of full-length transcripts than wild type M-MLV Reverse Transcriptase.

Features

•          Weak RNase H activity
•          High cDNA yield

Application

Synthesis of the first chain cDNA Library construction one-step RT-PCR primer extension 3′ and 5′ RACE.

Source

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

Unit definition

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

Quality control

1. Absence of Endonuclease
   • 1μg of Lambda DNA is incubated with 200 units of MMLV Reverse Transcriptase, RNase H Minus, 1×RT Reaction Buffer for 16 hour at 37°C. Following incubation, Lambda DNA is visualized as intact on an ethidium bromide-stained agarose gel to verify the absence of visible Endonuclease.
2. Absence of Nickase
   • 1μg of Type I supercoiled pBR322 is incubated with 200 units of MMLV Reverse Transcriptase, RNase H Minus, 1×RT Reaction Buffer for 16 hour at 37°C. Following incubation, the supercoiled DNA is visualized on an ethidium bromide-stained agarose gel to verify the absence of visible nicking or cutting.
3. Absence of Exonuclease
   • 1μg of Lambda DNA / Hind III Markers is incubated with 200 units of MMLV Reverse Transcriptase, RNase H Minus, 1×RT Reaction Buffer for 16 hour at 37°C. Following incubation, Lambda DNA / Hind III Markers is separated by 1% agarose gel and stained with ethidium bromide. Markers remain as intact bands without smearing.
4. Absence of RNase
   • 1ug of RNA is incubated with 200 units of MMLV Reverse Transcriptase, RNase H Minus, 1×RT Reaction Buffer for 4 hour at 37°C. Following incubation, the RNA is visualized as intact band on an ethidium bromide-stained agarose gel to verify the absence of visible RNase.
5. Function Assay
   • First-Strand cDNA Synthesis 200 units of enzyme incubated with 2.5ug of 8.3kb RNA for 1 hours at 42°C must synthesize ≥65% cDNA. ≥45% of the cDNA must be >6kb when analyzed by agarose gel electrophoresis.
6. Physical Purity
   • The purity is ≥95% as judged by SDS-polyacrylamide gel with Coomassie blue staining.

Product Number: IIA16

Storage condition

-20°C

Product component

Description

         HiFi II MMLV (H-) is a reverse transcriptase that uses engineered Escherichia coli to recombine and express mutated MMLV gene. This enzyme can catalyze complementary DNA polymerization using RNA or DNA: RNA hybrid chain as template. The mutated HiFi II MMLV (H-) reverse transcriptase RNase H activity is missing, reducing RNA degradation in reverse transcription reactions and making it easier to obtain full-length cDNA. HiFi II MMLV (H-) reverse transcriptase can synthesize the first strand of cDNA at 55 °C, providing higher specificity and stability. It can synthesize up to 12 kb of cDNA with high cDNA yield. Suitable for the synthesis of first strand cDNA, RT PCR, RT qPCR, and the construction of full-length cDNA libraries. The enzyme has been optimized with a unique A16 modification. IIA16 is a customized MMLV Reverse Transcriptase.

Unit definition

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

Quality control

200 U of this enzyme reacted with 1 µg of 16S, 23S rRNA at 37°C for 1 hour, and the electrophoretic bands of the RNA did not change

Note

• 1. 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 a dedicated area, using specialized instruments and consumables. Operators should wear masks and disposable gloves, and frequently change gloves.
• 2. 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℃ for 12 hours and sterilized under high pressure at 120℃ for 30 minutes before use. Alternatively, glass containers should be sterilized under dry heat at 180℃ 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.
• 3. 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.
• 4. If the initial amount of RNA is less than 50 ng, it is recommended to add RNA enzyme inhibitors.

Description

MMLV Reverse Transcriptase, encoded by Moloney Murine Leukemia Virus (MMLV 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. The difference between this to the general M-MLV RT is that the capacity to endure the heat is enhanced. It can remain the 100% activity at 50°C, it can also keep more than 80% activity even at 55°C.

Source

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

Applications

The first-strand cDNA synthesis; RT-PCR

Features

• lack RNase H activity: Weak RNase H activity High cDNA yield, can get more full length cDNA.
• thermoStable: the optimum reaction temperature is 50°C, the highest is 60°C. Can overcome the template RNA secondary structure, and finish the reverse transcriptase experiment smoothly.
• wide temperature range: can reverse transcript from 37-60C, with more than 80% of the highest activity at 42°C-55°C customer can choose the reaction temperature freely.
• 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.

Unit Definition

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

Concentration: 200U/μl

Package: Bulk

Component: M-MLV (200U/μl) 5xBuffer (with DTT)

Storage: -20°C

Order

The Lyophilized MMLV Reverse Transcriptase uses advanced lyophilization technology made to lyophilized powder format. The lyophilized MMLV can ship and keep at room temperature, it is very stable under RT without any activity lost. No need ship with dry ice, ice bag and other cold chain. Save much shipping cost, also no need to worry about the shipment delay cause the enzyme change to bad.

The Lyophilized MMLV Reverse Transcriptase is also a thermal stable reverse transcriptase. The thermal MMLV reverse transcriptase is recombined and expressed by mutant M-MLV gene using E. coli engineered bacteria. Thermal stable MMLV reverse transcriptase can be used in cDNA synthesis with RNA or RNA: DNA hybrids as templates. Loss of RNase H activity of Thermal stable MMLV reverse transcriptase reduces the degradation of RNA templates and makes it easier to obtain full-length cDNA. Thermal stable MMLV reverse transcriptase can synthesize the first strand of cDNA at 55°C, with higher specificity and stability, and can synthesize up to 12kb cDNA, with high cDNA yield. Thermal stable MMLV reverse transcriptase is suitable for Synthesis of first-strand cDNA, RT-PCR, RT-qPCR and the construction of full-length cDNA library, etc.

Feature

• Lyophilized format, convenience for transportation
• Thermal stable
• High Specificity
• High Sensitivity

Component

Storage

Store at room temperature.

Order