Reagent Center

PCR Kits

  • BioEasy Master Mix( SYBR Green, High ROX) MORE
    Product Details:

        Adopting nucleic acid amplification technique and combining intermediate fluorescence method, SYBR Green I real-time PCR can do detection and quantification on target gene fast and properly.

        The kit offers 2×SYBR Green Mix, including High ROX and other components except for template DNA and primers. During operation, user can do real time PCR reaction directly by adding primers and template DNA, which is very simple. The kit has the advantage of high efficiency in amplification, specificity, good repeatability, and high sensitivity and so on after the improvement of the reaction solution and the application of higher quality enzymes, and it’s high GC sequence reaction and quantify is greatly increased.

    Cat#

    BSB25L1B

    Components

    200Tests

     2×SYBR Green Mix(High ROX)

     1250 ul×4

    Instrument

    The reagent can be used in series Real Time PCR detection system of Bioer or the same kinds of the instruments of other companies;Especially suitable for Applied Biosystems 5700, 7000, 7300, 7700, 7900, 7900HT, 7900HT Fast; StepOne™, StepOne Plus™.

    Product Features:

    Using of the kit

    • 1  Upside down and mix before using the reagent. Before preparing PCR reagents, please centrifuge the reagent for a few seconds. Please mix, but the speed is less than 1400 rpm.
    • 2  Reaction mixture for real time PCR. (Work in ice).
    • 3  Fully mixing the reaction solution(50ul reaction), according to 45ul to each 0.2ml PCR reaction tube. Please add 5ul template in each PCR reaction tube. Adding template volume can be adjusted as appropriate but should not be exceeded 10ul.

    Note:

    • Please read this manual carefully before beginning the experiment.
    • For research use only.
    • Keeps all solution containing the fluorescence dye protected from light.
    • Nucleic acid is easy to be contaminated and resulted in to the untrusted result, so the user should strictly do the amplification experiment according to standard PCR process.

    Reagent storage

    This reagent can be stored at -20℃ and need be protected from light in 12 months. The reagent can be stored at 2-8℃ when you use up it in 2 weeks.

    Experimental data:

    Result analysis and judgments

    • 1   Input the concentrations of four positive References. Select fit point method to analyze. Confirm the base line (zero adjustment) by getting the fluorescent signals. Make the noise limit just beyond the peak of the amplification curve (rule less noise line) of normal negative control; then do quantitative analysis. You could also adjust by yourself according to the condition of instrument’s noises.
    • 2   Please record the concentration or Ct value of unknown sample after analysis.
    • 3   There are auto and manual for users choosing in melting curve analysis and record Tm value after analysis.
    • 4   Users also can use agarose gel electrophoresis to analyze specificity of PCR produce.
    Manual download
  • BioEasy Master Mix( SYBR Green, Low ROX) MORE
    Product Details:

    Description

        Adopting nucleic acid amplification technique and combining intermediate fluorescence method, SYBR Green I real-time PCR can do detection and quantification on target gene fast and properly.

        The kit offers 2×SYBR Green Mix, including Low ROX and other components except for template DNA and primers. During operation, user can do real time PCR reaction directly by adding primers and template DNA,which is very simple. The kit has the advantage of high efficiency in amplification, specificity, good repeatability, and high sensitivity and so on after the improvement of the reaction solution and the application of higher quality enzymes, and it’s high GC sequence reaction and quantify is greatly increased.

    Component   

    Cat#

    BSB25L1C

    Components

    200Tests

    2×SYBR Green Mix(Low ROX)

    1250 ul×4

    Instrument

    The reagent can be used in series Real Time PCR detection system of Bioer or the same kinds of the instruments of other companies; especially suitable for Applied Biosystems 7500, 7500 Fast, ViiA™7; Stratagene MX4000™, MX3005P™, MX3000P™, QuantiStudio™ 12K Flex .

    Product Features:

    Using of the kit

    • 1  Upside down and mix before using the reagent. Before preparing PCR reagents, please centrifuge the reagent for a few seconds. Please mix ,but the speed is less than 1400 rpm.
    • 2  Reaction mixture for real time PCR. (Work in ice).
    • 3  Fully mixing the reaction solution(50ul reaction), according to 45ul to each 0.2ml PCR reaction tube. Please add 5ul template in each PCR reaction tube. Adding template volume can be adjusted as appropriate but should not be exceeded 10ul.

    Reagent storage

    This reagent can be stored at -20℃ and need be protected from light in 12 months. The reagent can be stored at 2-8℃ when you use up it in 2 weeks.

    Note:

    • Please read this manual carefully before beginning the experiment.
    • For research use only.
    • Keeps all solution containing the fluorescence dye protected from sunlight.
    • Nucleic acid is easy to be contaminated and resulted in to the untrusted result, so the user should strictly do the amplification experiment according to standard PCR process.
    Experimental data:

    Result analysis and judgments

    • 1   Input the concentrations of four positive References. Select fit point method to analyze. Confirm the base line (zero adjustment) by getting the fluorescent signals. Make the noise limit just beyond the peak of the amplification curve (rule less noise line) of normal negative control; then do quantitative analysis. You could also adjust by yourself according to the condition of instrument’s noises.
    • 2   Please record the concentration or Ct value of unknown sample after analysis.
    • 3   There are auto and manual for users choosing in melting curve analysis and record Tm value after analysis.
    • 4   Users also can use agarose gel electrophoresis to analyze specificity of PCR produce.
    Manual download
  • BioEasy SYBR Green I Real Time PCR Kit MORE
    Product Details:

    Description

        Adopting nucleic acid amplification technique and combining intermediate fluorescence method, SYBR Green I real-time PCR can do detection and quantification on target gene fast and properly. The kit offers 2×SYBR Mix, including all components except for Taq DNA polymerase and primers. During operation, user can do real-time PCR reaction directly by adding primer, template and Taq DNA polymerase, which is very simple.

        SYBR Green I is one kind of fluorescence dye, which will excited fluorescence signal after specially inserted in dsDNA molecule. In PCR reaction system, when SYBR Green I dye combine with dsDNA, and excited with excited light, via testing strength of fluorescence, user can calculate the quantity of dsDNA. The advantage of fluorescence dye is that: it can be used in testing any dsDNA amplification, without design of probe, and make the testing method easier, meanwhile, it can decrease cost, what’s more, user can analyze melt curve directly. As soon as the PCR test finish. By the analysis of melt curve, user can judge whether there are existing aberrance or non-specific amplification. The Max Absorb wave length of SYBR Green I is 497nm, the Max. Emission wavelength is about 520nm.

    Component    

    Cat#

    BSB03M1

    BSB03L1

    Components

    100Tests

    200Tests

    2×SYBR Mix

    (with 4.0mM Mg2+)

    1250 ul×2

    1250 ul×4

    Taq DNA Polymerase

    32 ul

    32 ul×2

    25mM MgCl2

    1000 ul

    1000 ul×2

    ddH2O

    1000 ul×2

    1000 ul×4

    ROX Reference (50×)

    100 ul

    200 ul

    Instrument

    Bioer Line-Gene Real Time PCR Detection System or the same kind of the instruments in other companies.

    Product Features:

    Note:

    • 1. Please read this manual carefully before beginning the experiment.
    • 2. Don’t touch hand directly when you use one-time consumable such as pipe and centrifuge tube.
    • 3. Be caution during the whole process to ensure the accuracy of the experiment. After finishing the experiment, in order to avoid pollution, please use 75% alcohol to clear the worktable.
    • 4. The melted reagent should be stored as shorten as possible, and store them in environment of -20℃ immediately after testing.
    • 5. Please store reagent or PCR reaction system in shadow place due to the fluorescence dye contained
    • 6. Nucleic acid is easy to be contaminated and resulted in to the trusted result, so user should strictly do the amplification experiment according to standard PCR process.

    Sample storage

    Please store the sample at -20℃ and avoid repeating frozen.

    Experimental data:

    Result analysis and judgments

    • 1. Input the concentrations of four positive References. Select fit point method to analyze. Confirm the base line (zero adjustment) by getting the fluorescent signals. Make the noise limit just beyond the peak of the amplification curve (rule less noise line) of normal negative control; then do quantitative analysis. You could also adjust by yourself according to the condition of instrument’s noises.
    • 2. Please record the concentration or CT value of non-known sample after analysis.
    • 3. There are auto and manual for users choosing in melting curve analysis and record Tm value after analysis.
    • 4.  Users also can use agarose gel electrophoresis to analyze specificity of PCR products
    Manual download
  • BioEasy Taqman PCR Magic Mix MORE
    Product Details:

    Cat NO. BSB08L1

        This product is a standard reagent that is designed for the technology of Taqman probe. User can do real-time PCR reaction directly by adding primer, template and Taqman probe.

    Components

    Components

    Volume/ul

    Quantity

    2×Taqman Mix(with 6.0mM Mg2+)

    1000

    4

    Taq polymerase

    32

    2

    MgCl2

    1000

    2

    ddH2O

    1000

    2

    Applied Instrument

    Line-Gene series Real-time PCR system from BIOER TECHNOLOGY CO., LTD. or the same type of instrument from other companies.

    Product Features:

    Important Parameters

    • 1. Template

    This kit applies to plasmid DNA(10-107 copies), genomic DNA (100pg-1ug)or cDNA(1pg-100ng). To get the best result, the length of expansion segment should be in the bound of 50-200bp.

    • 2. Primer and Probe

    Primer and probe are the important parameters in the Real-time PCR reaction. During the time of designing primer and probe, suggest using special software, such as Primer Express, beacon Designer, AlleledID. Normally, the concentration of primer and probe is adjusting during 0.1uM-1.0uM.

    • Concentration of ion

    The PCR reaction liquid in this kit contains Mg2+ in end concentration 3mM. This is suitable for normal situation. However, you can adjust the concentration of Mg2+ for different intention segment. This kit contains an attaching tube of 25mM MgCl2.
    Adjust the concentration of Mg2+  according the following  list:

    End concentration of Mg2+(mM)

    Quantity of 25mM MgCl2 added into reaction(ul)

    4.0

    1.6

    5.0

    3.2

    6.0

    4.8

    Transport and storage condition

    Stored at -15℃~-20℃.

    Experimental data:

    Result analysis and determinant

    • 1. Input concentration of standard or frame of reference, select “Fit Points” method for analysis. Select a comparative smooth sect (without abnormal fluctuation) of fluorescence signal before the inflexion as base line adjustment. The threshold is the highest point where the base line does exceed normal negative contrast curve (ruleless noise line), and then do qPCR analysis. You may adjust according to instrument’s noise state.
    • 2. Record unknown sample’s concentration or CT value after analysis.
    • 3. Also could use gel electrophoresis to check and analyze the specificity of PCR product.

    Notices

    • 1. Please read this users’ manual carefully before doing experiment;
    • 2. This kit is just for research use, the result is just for reference;
    • 3. Please shorten the time of placing the reagents under room temperature as possible, and store them under -20℃ immediately;
    • 4. It’s possible to be polluted for nucleic acid expansion. So please accords to standard PCR operation instruction strictly.
    Manual download
  • BioRT cDNA First Strand Synthesis Kit MORE
    Product Details:

        BioRT cDNA First Strand Synthesis Kit provides the materials needed to rapidly and reliably synthesis first strand cDNA from RNA. It uses AMV reverse transcriptase to synthesize first strand cDNA from an RNA population.
        The kit includes all the necessary reagents for first strand cDNA synthesis. The kit includes both random primers and oligo(dT18) primers. The user can choose either of these or alternatively use gene specific primers. The reverse transcriptase in the BioRT cDNA First Strand Synthesis Kit is AMV, which provides up to 60℃ reverse transcriptase temperature and provides higher sensitivity and higher yield to cDNA’s synthesis and PCR. The kits can synthesis up to 10kb cDNA. The reaction buffer is optimized for a kind of reaction system.

    AMV Reverse Transcriptase(5U/μl)

    52 μl

    5 x RT Buffer

    500 μl

    dNTP Mixture(10mM)

    200 μl

    RNase inhibitor(40U/μl)

    52 μl

    Oligo-dT(18)

    52 μl

    Random Primer

    52 μl

    RNase free H2O

    1 ml

    Store at -20

    Product Features:

    RT Principle

    Notes
    • If using   random Hexamer Primer,  holding  at room  temperature for 10 minutes.
    • Reaction temperature can be elevated for RNA template with second structure, lower than 60℃.
    • Heat to 95℃ let AMV Reverse Transcriptase denature.

    Using Tips

    1.Total RNA or mRNA can be used as RNA template,and we suggest using Biozol(BSC51M1)to isolate high quality RNA;
    2. RNase contamination should be avoided and follow the measures:
    • Ware one time gloves and respirator because of the RNases in saliva and skin;
    • Use special instruments and consumables and handle in specific  areas;
    • Consumables should be treated at 180℃ for 60min or 37℃ for 12 hours with 0.1% DEPC H2O followed by sterilized at 121℃ for 30min;

    3. AMV reverse transcriptase, Taq polymerase and RNase inhibitor should be slowly pipetted after centrifuging, and put back at -20℃ later;
    4. Avoid frequently freezing and thawing dNTP;
    5.  Specific primers should be used and concentration of primers should be optimized and We suggest 0.4μM as a starting point for optimizing; Oligo-dT and Random primers are not suitable for this kit;

  • BioRT Master One Step RT-PCR Kit MORE
    Product Details:

    Description

    RT-PCR is a technology which combines reverse transcription (RT) and polymerase chain reaction (PCR) to detect target RNA. RT-PCR technology can be used to detect gene expression in cells and tissues, cDNA sequences of cloned specific genes and RNA viruses.
    Different from two steps RT-PCR in which RT and PCR separated in two tubes, BioRT Master One Step RT-PCR Kit is designed to combine RT and PCR in one tube, and no additional reagents are needed during the process.
    BioRT Master One Step RT-PCR Kit adopts one tube system, and this system can reduce the potential for contaminating the samples and provide specific, sensitive and convenient analysis of RNA.

    Cat#

    BSB07M2B

    Components

    100Tests

    2 × Master One-Step mix

    1250μl

    Enzyme mix

    125μl

    RNase free H2O

    1500μl

    RNA control(freeze dryness)

    20 μl

    RNA control S primer(5μM)

    10 μl

    RNA control A primer(5μM)

    10 μl

    Store at -20℃

    Product Features:

    One Step RT-PCR Principle

     

    Usage Tips

    1.   Total RNA or mRNA can be used as RNA template,and we suggest using Biozol(BSC51M1)to isolate high quality RNA;

    2.         Quality of primers affect the performance of BioRT Master One Step RT-PCR Kit; Factors such as GC percentage, length of primers and site of primers should be considered and we suggest primers be designed using special softwares.

    3.         RNase contamination should be avoided and follow the measures:

    1 Ware one time gloves and respirator because of the RNases in saliva and skin;

    2  Use special instruments and consumables and handle in specific areas;

    3 Consumables should be treated at 180for 2 hours or 37for 12 hours with 0.1% DEPC H2O followed by sterilized at 121for 30min;

    4.         Enzyme mix should be slowly pipetted after centrifuging, and put back at -20 later;

    5.         Specific primers should be used and concentration of primers should be optimized and We suggest 0.4μM as a starting point for optimizing; Oligo-dT and Random primers are not suitable for this kit.

    6.         Enzyme mix should be slowly pipetted after centrifuging, and put back at -20 later;

    7.         Specific primers should be used and concentration of primers should be optimized and We suggest 0.4μM as a starting point for optimizing; Oligo-dT and Random primers are not suitable for this kit.

     

    Usage Tips

    1.   Total RNA or mRNA can be used as RNA template,and we suggest using Biozol(BSC51M1)to isolate high quality RNA;

    2.         Quality of primers affect the performance of BioRT Master One Step RT-PCR Kit; Factors such as GC percentage, length of primers and site of primers should be considered and we suggest primers be designed using special softwares.

    3.         RNase contamination should be avoided and follow the measures:

    1 Ware one time gloves and respirator because of the RNases in saliva and skin; 2  Use special instruments and consumables and handle in specific areas;

    3 Consumables should be treated at 180for 2 hours or 37for 12 hours with 0.1% DEPC   H2O followed by sterilized at 121for 30min;

    4.         Enzyme mix should be slowly pipetted after centrifuging, and put back at -20 later;

    5.         Specific primers should be used and concentration of primers should be optimized and We suggest 0.4μM as a starting point for optimizing; Oligo-dT and Random primers are not suitable for this kit.

  • BioRT One Step RT-PCR Kit MORE
    Product Details:
    Description

        RT-PCR is a technology which combines reverse transcription (RT)  and polymerase chain reaction (PCR) to detect target RNA. RT-PCR technology can be used to detect gene expression in cells and tissues, cDNA sequences of cloned specific genes and RNA viruses.
        Different from two steps RT-PCR in which RT and PCR separated in two tubes, BioRT One Step RT-PCR Kit is designed to combine RT and PCR in one tube, and no additional reagents are needed during the process. High-quality AMV reverse transcriptase produced in USA and Hot start Taq polymerase are included in this kit, and an optimized buffer ensure the best performance of the two enzymes. AMV reverse transcriptase that produce high-yield cDNA and highly purified recombinant Taq polymerase with monoclonal antibody used together can get up to 4kb products.
        BioRT One Step RT-PCR Kit adopts one tube system, and this system can reduce the potential for contaminating the samples and provide specific, sensitive and convenient analysis of RNA.

    Cat#

    BSB07M1

    Components

    100Tests

    AMV Reverse Transcriptase(5U/μl)

    50μl

    Taq polymerase(5U/μl)

    50μl

    10X RT-PCR Buffer(30mM Mg2+ plus)

    500μl

    dNTP Mixture(2.5mM)

    500μl

    MgCl2(25mM)

    500μl

    RNase inhibitor(40U/μl)

    100μl

    RNase free H2O

    100μl×2

    RNA control(freeze dryness)

    20 μl*

    RNA control S primer(5μM)

    20 μl

    RNA control A primer(5μM)

    20 μl

    Store at -20℃

    Product Features:
    One Step RT-PCR Principle

    Usage Tips

    1. Total RNA or mRNA can be used as RNA template ,and we suggest using Biozol (BSC51M1) to isolate high quality RNA;

    2. Quality of primers affect the performance of BioRT One Step RT-PCR Kit; Factors such as GC percentage, length of primers and site of primers should be considered and we suggest primers be designed using special softwares.

    3. RNase contamination should be avoided and follow the measures:

    • Ware one time gloves and respirator because of the RNases in saliva and skin;
    • Use special instruments and consumables and handle in specific areas;
    • Consumables should be treated at 180℃ for 2 hours or 37℃ for 12 hours with  0.1% DEPC H2O followed by sterilized at 121℃ for 30min;

    4. AMV reverse transcriptase, Hot Start Taq polymerase and RNase inhibitor should be slowly pipetted after centrifuging, and put back at -20℃ later;

    5. Avoid frequently freezing and thawing dNTP;

    6. Specific primers should be used and concentration of primers should be optimized and We suggest 0.4μM as a starting point for optimizing; Oligo-dT and Random primers are not suitable for this kit;

    7. Concentration of MgCl2 can be optimized and 0.5mM/time is suggested when the length of target template is longer than 2kb;

  • BioRT Real Time RT-PCR Kit(SYBR Green) MORE
    Product Details:

    Introduce

        This product is a 2 × Master Mix for “One-step Real-time PCR” using a thermostable DNA polymerase. This system allows for “One-step Real-time PCR”, including reverse transcription and PCR steps. This reagent is applicable for SYBR Green assay.

        This product adopts one tube system, and this system can reduce the potential for contaminating the samples and provide specific, sensitive and convenient analysis of RNA.

    Cat#

    BSB33S1

    BSB33M1

    Components

    50Tests

    100Tests

    RT-PCR Mix (SYBR Green)

    750 ul

    750 ul×2

    Enhancer

    75 ul

    150 ul

    Storage: -20℃

    Product Features:

        Choose F1(SYBR Green I) channels when collecting fluorescent signals and set fluorescent signals detecting at 72 ℃ . Before running Line-Gene Series Real-time PCR detection system, set fluorescent signals detecting at 72℃ for 5 seconds, and then adjust gain to background between 5-15.

         NoteAnnealing temperature can adjust by primer annealing temperature.

    Usage Tips

    1. Total RNA or mRNA can be used as RNA template,and we suggest using high quality RNA;
    2. Quality of primers affect the performance of BioRT Real Time RT-PCR Kit (SYBR Green); Factors such as GC percentage, length of primers and site of primers should be considered and we suggest primers be designed using special software.3.
    3. RNase contamination should be avoided and follow the measures:
    • Ware one time gloves and respirator because of the RNases in saliva and skin;
    • Use special instruments and consumables and handle in specific areas;
    • Consumables should be treated at 180℃ for 2 hours or 37℃ for 12 hours with 0.1% DEPC H2O followed by sterilized at 121℃ for 30min;
    4. RT-PCR Mix (SYBR Green) should be slowly pipetted after centrifuging, and put back at -20℃ later.
    Experimental data:

    Figure 1(Real-time PCR Curve):Detect GAPDH gene of total RNA dilution in the mouse(270ng、90ng、 9ng、0.9ng、0.09ng、0.03ng、0.01ng and negative).

    Figure 2(Standard Curve):Total RNA dilution in the mouse.

    Figure 3(melting curve):Total RNA dilution in the mouse.

    Manual download
  • BioRT Real Time RT-PCR Kit (With ROX) MORE
    Product Details:

    Introduce

         This product is a RT-PCR Reaction Solution for “one-step real-time PCR” using a thermostable DNA polymerase. This system allows for “one -step real-time PCR”, including reverse transcription and PCR steps. This reagent is applicable for TaqMan® assay or SYBGreen assay.
        This product adopts one tube system, and this system can reduce the potential for contaminating the samples and provide specific, sensitive and convenient analysis of RNA. RT-PCR reaction Solution containing ROX dye can be used for ROX calibration.

    Cat#

    BSB23M2B

    Components

    100Tests

    RT-PCR Reaction Solution (with ROX)

    750 ul×2

    Enhancer

    150 ul

    Stabilizer

    150 ul

    Product Features:
    Usage Tips
    • 1. Total RNA or mRNA can be used as RNA template,and we suggest using high quality RNA;
    • 2. Quality of primers affect the performance of BioRT One Step RT-PCR Kit; Factors such as GC percentage, length of primers and site of primers should be considered and we suggest primers be designed using special software.
    • 3. RNase contamination should be avoided and follow the measures:
    •  Ware one time gloves and respirator because of the RNases in saliva and skin;
    •  Use special instruments and consumables and handle in specific areas;
    •  Consumables should be treated at 180℃ for 2 hours or 37℃ for 12 hours with 0.1% DEPC H2O followed by sterilized at 121℃ for 30min;
    • 4. RT-PCR Reaction Solution should be slowly pipetted after centrifuging, and put back at -20℃ later.
  • BioRT Two Step RT-PCR Kit MORE
    Product Details:

        RT PCR is a useful technique for the investigation of gene expression, viral load, pathogen detection, and numerous other applications. When analyzing gene expression or viral load, the RNA of interest first needs to be reverse transcribed into cDNA. The subsequent PCR can be performed separately (two-step RT-PCR).

         BioRT Two Step RT-PCR Kit is designed for two step RT-PCR of RNA samples from various  sources.The kit includes all the necessary reagents for cDNA synthesis and subsequent PCR. Either total RNA, messenger RNA, viral RNA or in vitro transcribed RNA can be used as a template for reverse transcription. The kit includes both random primers and oligo(dT18) primers. The user can choose either of these or alternatively use gene specific primers. The  reverse transcriptase in  the BioRT Two  Step RT-PCR  Kit is AMV,  which provides up to 60℃ RT temperature and provides higher sensitivity and higher yield to cDNA’s synthesis and PCR. The performance of the PCR step is based on a high fidelity Taq mix DNA polymerase. The kits can synthesis up to 12kb cDNA and 6kb DNA. The reaction buffer is optimized for a kind of reaction system (10ul for RT and 25ul for PCR).

    Cat#

    BSB05M1

    Components

    100Tests

    AMV Reverse Transcriptase(5U/μl)

    52 μl

    5 x RT Buffer

    500 μl

    dNTP Mixture(10mM)

    52 μl

    RNase inhibitor(40U/μl)

    52 μl

    Oligo-dT(18)

    52 μl

    Random Primer

    52 μl

    RNA control (freeze dryness)

    Add 20 μl *

    RNase free H2O

    1 ml×2

    Taq mix DNA polymerase(5U/μl)

    52 μl

    10 x PCR Buffer (include 15mM Mg2+)

    500 μl

    MgCl2(25mM)

    200 μl

    RNA control S primer   (5μM)

    20 μl

    RNA control A primer   (5μM)

    20 μl

    Product Features:

    RT-PCR Principle

    Note:
    • RT products can up to 5.0 μl.
    • 5℃  lower than Tm value of primers;Control primer is 55℃.
    • Add to 40-45 cycles when detecting rare RNA templates.
    • Usually 1kb/min. Control primer is 45s.

    Using Tips

    • 1. Total RNA or mRNA can be used as RNA template,and we suggest using Biozol(BSC51M1)to isolate high quality RNA;
    • 2. RNase contamination should be avoided and follow the measures:
    • Ware one time gloves and respirator because of the RNases in saliva and skin;
    • Use special instruments and consumables and handle in specific areas;
    • Consumables should be treated at 180℃ for 60min or 37℃ for 12 hours with 0.1% DEPC H2O followed by sterilized at 121℃ for 30min;
    • 3. AMV reverse transcriptase, Taq polymerase and RNase inhibitor should be slowly pipetted after centrifuging, and put back at -20 later;
    • 4. Avoid frequently freezing and thawing dNTP;

    • 5. Concentration of MgCl2 can be optimized and increasing 0.5mM/time is suggested when the length of target template is longer than 2kb;

    • 6. Quality of primers affect the performance of BioRT One Step RT-PCR Kit; Factors such as GC percentage, length of primers and site of primers should be considered and we suggest primers be designed using special soft wares.

    1.                AMV reverse transcriptase, Taq polymerase and RNase inhibitor should be slowly pipetted after centrifuging, and put back at -20 later;

    2.                Avoid frequently freezing and thawing dNTP;

     

    3.                Specific primers should be used and concentration of primers should be optimized and We suggest 0.4μM as a starting point for optimizing; Oligo-dT and Random primers are not suitable for this kit;

    4.                Concentration of MgCl2 can be optimized and increasing 0.5mM/time is suggested when the length of target template is longer than 2kb;

    5.                Quality of primers affect the performance of BioRT One Step RT-PCR Kit; Factors such as GC percentage, length of primers and site of primers should be considered and we suggest primers be designed using special soft wares.

    2
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