Reagent Center

General Reagent

  • BioReady LA Taq MORE
    Product Details:

    Introduction

        BioReady LA Taq is a thermostable DNA polymerase with 3′ → 5′ exonuclease activity. It has the characteristic of higher efficiency and higher fidelity. This enzyme is basically applicable to different kinds of template DNA. It can amplify the simple template DNA up to 30 kbp fragments effectively. And it can amplify the template DNA having complex secondary structure up to 12 kbp fragments effectively. PCR products amplified an “A” base at 3’end by using the enzyme. The product is suitable for cloning into TA vector system.

    Component

    Cat#

    BSA13S2

    BSA13M2

    Components

    250U

    500U

    10 ´ Reaction Buffer (with 15mM MgCl2)

    1ml

    1ml×2

    BioReady LA Taq

    (5U/μl)

    50μl

    100μl

    25mM MgCl2

    500 ul

    1000 ul

    6 ´ Loading Dye

    500 ul

    500 ul

    Storage

    -20℃。

    Product Features:

    Applications

        BioReady LA Taq is optimized for high fidelity amplification of long template DNA or templates DNA having complex secondary structure, which can be used in construction of gene mapping, sequencing and other research on molecular genetics.

    Purity

    1)   Endonuclease and exonuclease activity are not detected after the incubation of 0.6 µg of supercoiled λDNA-Hind III with 10 units of this enzyme for 1 hour at 74°C.
    2)   Endonuclease and exonuclease activity are not detected after the incubation of 0.6 µg of supercoiled Supercoiled pBR322 DNA with 10 units of this enzyme for 1 hour at 74°C.
    3)   Endonuclease and exonuclease activity are not detected after the incubation of 0.6 µg of supercoiled λDNA with 10 units of this enzyme for 1 hour at 74°C.

  • BioReady Pfu pac. MORE
    Product Details:

    Description:

        BioReady Pfu is a thermostable enzyme of approximately 92 kDa isolated from pyrococcus furiosus. BioReady Pfu catalyzes the DNA-dependent polymerization of nucleotide into duplex DNA in the 5’3’ direction in the presence of magnesium ions. The enzyme also exhibits 3’5’exonuclease (proofreading) activity. Base misinsertions that may occur infrequently during polymerization are rapidly excised by the proofreading activity of the polymerase. Consequently, BioReady Pfu is useful for polymerization  reactions requiring high fidelity synthesis.

    Kit Components

    Cat#

    BSA10S1

    BSA10M1

    Components

    100units

    250units

    10 ´ Reaction Buffer (with 20mM MgSO4)

    400μl

    1000μl

    BioReady Pfu(5U/μl)

    80 μl

    200 μl

    2.5mM dNTP Mixture

    300μl

    500μl

    6×loading dye

    200μl

    500μl

    ddH2O

    1250μl

    1250μl

     

    Product Features:

    Note:

    • It is critical to withhold BioReady Pfu until the addition of dNTP; otherwise, the proofreading activity of the polymerase may degrade the primers, resulting in nonspecific amplification and reduced product yield. Assemble components on ice.
    • If using a thermal cycler without hot lid, overlay the reaction mix with 1-2 drops (approximately 50μl) of the mineral oil to prevent evaporation during thermal cycling. Centrifuge the reaction mix in a micro centrifuge for 5 seconds.
    •  Immediately place the reactions in a thermal cycler that has been preheated to 95℃ for 1-2 minutes to ensure that the target DNA is completely denatured. Incubation for longer than 2 minutes at 95℃ is unnecessary and may reduce the yield due to DNA damage.
    • Start the thermal cycling program. The cycling profile given in Table 1 may be used as a guideline. Optimize the amplification profile for each primer target combination.

    General Consideration

    Enzyme consideration
    We recommend that 1.25 units of BioReady Pfu be used per 50μl amplification reaction. The inclusion of more enzyme will increase 3’5’exonuclease (proofreading) activity. It is essential to withhold BioReady Pfu from the reaction until after the addition of the dNTP mixture and to assemble components on ice.
    Primer design
    The sequences of the primers are a major consideration in determining the optimal temperature of the PCR amplification cycles. For primers with a high Tm, it may be advantageous to increase the annealing temperature. Higher temperature minimized nonspecific primer annealing, increase the amount of specific product and reduces the amount of primer-dimer formation.

    The 3’5’ exonuclease activity may degrade primers. To overcome the degradation, longer primers with maximized GC content could be used. Primers can also be protected by introducing phosphothioate bonds at their 3’ termini.
     Extension time
    The extension rate of BioReady Pfu is lower than of Taq DNA polymerase. Therefore, during the extension step, allow approximately 2 minutes for every 1kb to be amplified (minimum extension time of 1 minute). For most reaction, 25-35 cycles are sufficient.

     

  • BioReady Taq Mix(with dye) MORE
    Product Details:
    Introduction

         This product has 94kD thermostable DNA polymerase. Principle-based enzyme kinetics of genetic engineering in a special mutation so that the enzyme activator to reduce the dependence of the enzyme amplified greatly improves efficiency, reduces the rate mismatch. PCR produce at 3’end has an “A” base amplified by the use of the enzyme. The produce is suitable for cloning by TA vector system.
        This product has 2×Taq Mix  (with dye)and ddH2O. Only adding primer and the template can be amplified on the machine, and that this product has good stability, amplification efficiency, specificity and other advantages.

    Cat#

    BSA31S1B

    BSA31M1B

    Components

    50Tests

    200Tests

    2×Taq Mix(with dye)

    1250 μl

    5000 μl

    ddH2O

    1000 μl

    4000 μl

    Product Features:

    Applications

     This product is small in the rate mismatch, suitable for large-scale genetic testing. This product is suitable for less than 8kb DNA, DNA markers, DNA sequencing, and its amplified produce can be directly used for TA cloning. Using is very quick and easy.

    Reagent Storage

     Please store the reagent at 2-8℃. It cannot store at -20℃,otherwise its quality will decline.

    For research use only.

  • BioReady Taq Mix MORE
    Product Details:

    Introduction

         This product has 94kD thermostable DNA polymerase. Principle-based enzyme kinetics of genetic engineering in a special mutation so that the enzyme activator to reduce the dependence of the enzyme amplified greatly improves efficiency, reduces the rate mismatch. PCR produce at 3’end has an “A” base amplified by the use of the enzyme. The produce is suitable for cloning by TA vector system.

        This product has 2×Taq Mix and ddH2O. Only adding primer and the template can be amplified o the machine,and that this product has good stability, amplification efficiency, specificity and other advantages.

    Cat#

    BSA31S1

    BSA31M1

    Components

    50Tests

    200Tests

    2×Taq Mix

    1250 μl

    5000 μl

    ddH2O

    1250 μl

    5000 μl

    Product Features:

    Applications

    This product is small in the rate mismatch, suitable for large-scale genetic testing. This product is suitable for less than 6kb DNA, DNA markers, DNA sequencing, and its amplified produce can be directly used for TA cloning. Using is very quick and easy.

    Reagent Storage

     Please store the reagent at 2-8℃. It cannot store at -20℃,otherwise its quality will decline. 

    For research use only.

  • BioReady Taq pac. MORE
    Product Details:

    Description

        BioReady Taq is a highly pure, thermostable recombinant DNA polymerase encoded by a modified  gene  from  Thermus  aquaticus  species  and  expressed  in  E.coli.  Its recombinant nature ensures utmost purity, reproducibility and processivity. BioReady Taq processes an initial 5'’3’ exonuclease activity and lacks 3’5’exonuclease activity. The enzyme leaves a single 3’-nucleotide overhang that make the products suitable for cloning by TA vector system.

        BioReady Taq provides a thermostability that meets the requirements of specialized PCR applications.

    Cat#

    BSA09S2

    BSA09M2

    Components

    200units

    500units

    10 ´ Reaction Buffer (with 15mM MgCl2)

    500μl

    1000μl

    BioReady Taq (5U/μl)

    100μl

    200μl

    2.5mM dNTP Mixture

    350μl

    800μl

    25mM MgCl2

    500μl

    500μl

    6×loading dye

    300μl

    500μl

    ddH2O

    1250μl

    1250μl×2

     

    Product Features:

    Physical purity

    BioReady Taq is determined to be>90% pure as judged by SDS-PAGE gels with Coomassie brilliant blue staining.

    Endonuclease Assay

    One microgram of super coiled plasmid DNA (pUC18) is incubated with 5 units of BioReady Taq for 8 hours at 45℃ and 8 hours at 70℃ in 1´ reaction buffer. Following incubation, the DNA is visualized as a single band an ethidium bromide-stained agarose gel.

    Exonuclease Assay

    One microgram of lambda DNA and 1ug lambda/HindIII DNA are incubated with 5 units of BioReady Taq for 8 hours at 45℃and 8 hours at 70℃ in 1´ reaction buffer. Following incubation, the DNA is visualized as a single band an ethidium bromide-stained agarose gel.

    Functional Assay

    BioReady Taq is tested for performance in the polymerase chain reaction using 2.5 units of enzyme to amplify a lambda DNA (0.5kb, 1.0kb and 3kb) and beta-globin partial fragment (408bp) of human. The resulting PCR product is visualized as a single band an ethidium bromide –stained agarose gel.

    Unit Definition

    One unit is defined as the amount of enzyme that incorporates 10 nmoles of dNTPs into acid-insoluble form within 30 min at 72℃ TAPS assay condition.

    Storage buffer

    20 mM Tris-HCl (pH8.0 at 25℃), 100 mM KCl ,0.1mM DDT, 50% Glycerol, 0.5% Tween 20 and 0.5%NP-40.

    Reaction buffer: 10X Reaction buffer with magnesium [proprietary formulation]

    Applications

    Routine PCR, TA cloning, RT-PCR, Multiplex PCR, PCR-SSCP, PCR-RFLP and PCR-RAPD, etc.

    Storage temperature

    Store at -20℃. Avoid exposure to frequent temperature changes.

     

  • Red Blood Cell Lysis Buffer MORE
    Product Details:

    Introduction

         Red Blood Cell Lysis Buffer has been developed for the preferential lysis of red blood cells from human whole blood. The buffer provides a very simple, fast and economic way for isolating white cells from whole blood without destroying white cells. In generally, we can get 80% or more white cells from the sample.

    Kit Components

    Cat#

    BSA06M1

    Red Blood Cell Lysis Buffer

    120ml

    Handbook

     1 copy

    Product Features:

    Storage

    1. The solution is to be stored at 2-8℃.
    2.  The solution is stable for 24 months from the date of manufacture.

    Important Notes

    Do

    Do not

    Warm the buffer to +15 to +25°C prior to use.

    Do not use blood that has been frozen and thawed more than three times for any DNA Applications. Warm blood to +15 to +25°C.

    Perform all centrifugation steps at +15 to +25°C in a variable-speed microfuge.

    Do not use blood that has been stored for longer than one month for any DNA applications.

    Use blood stored in EDTA, citrate, or heparin anticoagulants.

    The buffer is intended for the removal of red blood cells from human blood, may not be appropriate for the lysis of red blood cells of animals.

    Use blood that has been stored for ≤1 month at +2 to

    +8°C.

     

    For best results in DNA applications, use fresh  blood or blood stored for ≤7 days.

     

        In addition, be certain to follow all universal safety precautions governing work with biohazardous materials. For example, wear lab coats, gloves, and safety glasses at all times. Also, properly dispose of all contaminated materials, decontaminate work surfaces, and use a biosafety cabinet whenever aerosols might be generated.

    Application

        Red Blood Cell Lysis Buffer can be used to lyse red blood cells from 1µl ~10ml human whole blood, yielding intact white blood cells for future applications. This buffer is not intended for use with whole blood from any other species.

  • RT007 AMV Reverse Transcriptase MORE
    Product Details:

    Description

        RT007 AMV reverse transcriptase was isolated from Avian Myeloblastosis Virus (AMV) as the ab holoenzyme of molecular weight 157,000 Daltons, using a modification of the method described by Houts et al. (1). This preparation is essentially free of nuclease. It is qualified for cDNA synthesis and also for dideoxy sequencing of DNA and RNA. Total RNA or poly A+ RNA can be used as template,and optimal temperature is 42℃-55℃, up to 60℃. If NaPPi is used, 37-41℃ is preferred. RT reaction buffer covered by patents can be used either as 5X /10X stock depending on different purpose (details in standard application). Up to 10-12kb cDNA may be obtained according to standard application.

    Cat#

    BSA01S2

    BSA01M2

    Components

    200 Units

    1000 Units

    RT Buffer(5X/10X)

     1.0ml

     1.0 ml×4

    AMV Reverse Transcriptase (10U/ml)

    20ml

    100ml

    Product Features:

    Storage conditions

    It is strongly suggested that the enzyme be aliquoted and stored at -70℃, although storage at -20℃ is adequate for short periods of time.

    Attention:

    1. Repeated freezing and thawing results in loss of enzyme activity.
    2. Do not store this enzyme in a frost-free freezer.
    3. Due to the viscosity of the enzyme, maximum dispensing efficiency is achieved by the use of positive displacement pipets.

    Quality Assurance Assay

    Ribonuclease Assay: Fifteen units of AMV RT were incubated with 1μg of RNA Ladder for two hours at 37℃, in 1´ reaction buffer. No more than the equivalent of 4´10-8 Unit of RNase 1A was detected. This assay is capable of detecting 2´10-8 Units of RNase 1A.

    Enzyme Storage Buffer

    200 mM

    Potassium Phosphate, pH 7.2

    2.0 mM

    Dithiothreitol(DTT)

    50%

    Glycerol(v/v)

    0.2%

    Triton X-100

  • BioReady rTaq MORE
    Product Details:

    Introduction

        This product is a 94kD thermostable DNA polymerase. Principle-based enzyme kinetics of genetic engineering in a special mutation so that the enzyme activator to reduce the dependence of the enzyme amplified greatly improves effciency, reduces the rate mismatch. PCR products amplified by the use of the enzyme 3’end with an “A” base. The product is suitable for cloning by TA vector system.

    Cat#

    BSA12S1

    BSA12M1

    Components

    250U

    500U

    10 ´ Reaction Buffer (with 15mM MgCl2)

    1 ml

    1 ml×2

    6 ´ loading dye

    250 μL

    500 μL

    BioReady rTaq

    (5U/ μL)

    50 μL

    100 μL

    Storage

    -20℃。

    Product Features:

    Applications

    PCR amplification of DNA fragments less than 6 kb, DNA labeling, DNA sequencing, Generation of PCR product for TA cloning.

    Result test

    Take 5 μl PCR products each tube for electrophoresis after PCR reaction.

    Quality Control

     

    • 1)5'– 3' exonuclease activity                           Yes
    • 2)Extra A addition                                             Yes
    • 3)3'–5' exonuclease activity                            No
    • 4)DNase contamination                                 No
    • 5)RNase contamination                                 No
    • 6)Protease contamination                              No
    • 7)DNA contamination                                      No
    • 8)RNA contamination                                      No

     

    This product was purified by HPLC for 2 times, various contamination was effectively eliminated. It keeps the enzyme high purity and high activity.

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