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Combo Includes:
200 μL Taq DNA Polymerase (5 units/μL)
4 x 500 μL vials of dNTP Premix (100 mM each of dATP, dCTP, dGTP, dTTP)

D118-1000

High-performance Taq DNA Polymerase specially purified to produce excellent yields with little or no background. Its outstanding activity and unique thermal DNA amplification properties make it one of the most economic DNA polymerases available.

Special Features

  • Robust performance
  • Leaves 3'A overhang
  • Stable at all storage temperatures
  • Best price/quality
  • APPLICATIONS
  • General PCR
  • Genomic analysis
  • TA cloning

D116

Ready-to-use, ultra-pure, molecular grade solution made from 100mM each of dATP, dCTP, dGTP, and dTTP stock. The PreMix is designed to reduce "hands-on" time for researchers, thereby lowering the possibility of contamination and error. Furthermore, every lot of dNTP solution is pre-tested for optimal performance. The nucleotides are supplied in sterile, doubly distilled water. Each dNTP has a final concentration of 10mM for a total PreMix dNTP concentration of 40mM.

Special Features

    • Ready to use
    • Each lot functionally tested
    • Purity; at least >99%
    • APPLICATIONS
    • For RT-PCR, large PCR, and real-time PCR
    • Reverse Transcription and mutagenesis
Item#:
ASCOMBOTAQ

Hotstart Taq 2x Mastermix,

5x1ml, 200x50ul rxns

2X PCRmaster mix with HotStart Taq DNA polymerase

High performance, ready-to-use 2X PCRmaster mix with HotStart Taq DNA polymerase, in red color. Users only need to add template, primers and H2O for the reaction. The HotStart PCR 2x Master Mix contains a inert, non-toxic red dye to visualize PCR mixing step, and also allows direct loading of PCR products on to gels  for electrophoresis.

The HotStart Taq DNA Polymerase is a chemically modified Taq DNAPolymerase, whose enzyme activities can only be activated after 3-5 minutes of incubation at 95°C. The HotStart Taq Polymerase uses amplification conditions for regular Taq DNA Polymerase, except no polymerase activity will be present before the onset of thermal cycling. This prevents nonspecific DNA amplification and primer dimer formation. The amplified products, up to 7Kb in length, contain PCR products with blunt and 3-overhanging A end.  This allows flexible protocols for PCR cloning, if the amplified fragments need to be cloned.

SPECIAL FEATURES

  • All-in-one 2X master mix format
  • Hot start
  • Pre-optimized
  • Direct gel loading
  • In green color
  • up to 7kb in length
  • Item#:
    D136
    Your Price:
    160.60
    Each

    Bullseye High Fidelity PCR Master Mix


    The newly developed Bullseye HFL PCR Master Mix represents the highest fidelity for a PCR product. Bullseye's HFL PCR Master Mix has eight times higher fidelity when compared to Pfu, previously considered to be the golden standard for high fidelity PCR. This Master Mix also works well for large PCR fragments. 

    The Bullseye HFL PCR Master Mix is in a 2X format and contains modified, high fidelity thermal stable DNA polymerases, in a pre-optimized PCR buffer. The amplified DNA will be blunt-ended. When using this master mix for most PCR experiments, only template, primers and H2O will be needed.

    SPECIAL FEATURES & APPLICATIONS
    • High fidelity with robustness
    • Large fragment PCR
    • 2X master mix - makes PCR easy and consistent
    • For cloning, promoter study, RACE, DNA sequencing and more
    Item#:
    BE135HFL
    Your Price:
    88.00
    Each

    Eco-Taq MasterMix, 2X,

    1mL, 1 vial of 1000ul

    Affordable price & shipped at room temperature.

     

    A premixed, ready-to-use solution for efficient amplification of DNA templates by PCR.

    • 2X Eco-Taq MasterMix contains Taq DNA Polymerase, dNTPs, Mg2+ and Reaction Buffer at optimal concentrations for efficient amplification of DNA templates by PCR, as well as an inert loading dye.
    • This pre-mixed formulation saves time and reduces contamination due to the fewer pipetting steps required for PCR set up. To prepare the final PCR, only primers and template DNA need to be added.
    • The mix retains all features of Taq DNA Polymerase and can amplify DNA targets up to 5 kb (simple template).
    • The elongation velocity is 0.9~1.2kb/min (70~75°C).
    • It has 5' to 3' polymerase activity but lacks 3' to 5' exonuclease activity which results in a 3'-dA overhang PCR product.

     

    Features:

    • Convenient: Just add primers and template DNA
    • High yields of PCR products with minimal optimization.
    • High efficiency: saves your time by simplifying the process
    • Reproducible: lower contamination risk and pipetting error.

      Applications:

    • High-throughput PCR.
    • Routine PCR with high reproducibility
    • Generation of PCR products for TA cloning

      Contents:
      2X Taq Mix 1ml

      Store at -20°C
      For research use only

    Item#:
    ECO-TAQ1
    Your Price:
    20.90
    Each

    Accuris™ Taq Plus

    • Higher fidelity for long PCR amplicons, up to 35kb
    • Ideal for problematic templates
    • Better sensitivity and higher activity for low copy and long PCR
    • Enzyme of choice for TA cloning

    Accuris Taq Plus, an optimized blend of our Hot Start Taq and a proofreading polymerase, provides 5x better fidelity than wild-type Taq and increased enzyme activity. It is the perfect choice for GC and AT rich templates, low copy number and long PCR.
    The 3' -5' exonuclease activity of the proofreading enzyme produces fragments suitable for TA cloning. Accuris Taq Plus is provided with a 5X buffer with dNTPs or in a convenient one tube 2X Master Mix.

     

    Item No.

    Description

    Volume

    PR1000-TP-S

    Accuris Taq Plus - Sample

    50 units

    PR1000-TP-250

    Accuris Taq Start Taq DNA Polymerase

    250u (5u/p1)

    PR1000-TP-500

    Accuris Taq Start Taq DNA Polymerase

    1000u (5u/p1)

    PR1001-TP-S

    Accuris Taq Plus Master Mix, 2X conc. - Sample

    20 reactions

    PR1001-TP-200

    Accuris 2X Taq Plus Master Mix

    200x 50µl Reactions

    PR1001-TP-1000

    Accuris 2X Taq Plus Master Mix

    1000x 50µl Reactions

    Item#:
    ASACCREAG3

    PR DNA Polymerase, High Fidelity, 2.5 U/µL

    • Provides higher fidelity than standard Taq DNA Polymerase
    • Produces blunt-ended fragments
    • Processes <3 kb with extremely high fidelity

     

    Cat. No.

    Units

    10X Ammonium Buffer (MgCl2
    15 mM)

    MgCl2
    25 mM

    BE211102

    250

    1.5 mL

    1.5 mL

    BE210303

    500

    1.5 mL

    1.5 mL

    BE211104

    1,000

    2 x 1.5 mL

    2 x 1.5 mL

    BE211106

    2,500

    4 x 1.5 mL

    4 x 1.5 mL

    Store at -20°C. For in-vitro laboratory use only

    Bullseye PREMIUM PR DNA Polymerase is a thermostable enzyme with proofreading ability, which can be used in primer extension reactions and other molecular biology applications. PR Polymerase exhibits both 5'-3' DNA polymerase activity and 3'-5' proofreading exonuclease activity. It is recommended for applications, which require extremely high fidelity or blunt ending.


    Optimal reaction conditions are achieved by using the 10x Ammonium buffer containing MgCl2 provided with the enzyme. 25 mM MgCl2 is also included separately, in case a higher MgCl2 concentration is required for a specific reaction.


    Unit Definition
    One unit is defined as the amount that incorporates 10 nmoles of dNTPs into acid-precipitable form in 30 minutes at 72°C under standard assay conditions.

    10X Ammonium Reaction Buffer
    Tris-HCl pH 8.5, (NH4)2SO4, 1% Tween20,
    15mM MgCl2

    PR Storage Buffer
    50 mM Tris-HCl (pH 8.0), 50 mM NaCl, 0.1 mM EDTA, 1 mM DTT, 50% Glycerol, 0.1% NP40, 0.1% Tween-20.

    Quality Control
    Endonuclease, exonuclease and priming activities are not detected after 3 hours incubation of 1 mg of pUC19 plasmid DNA and 0.5 µg EcoR I digested lambda phage DNA at 72°C in the presence of 40 units of PR DNA Polymerase.

    Suggested Protocol using PR DNA Polymerase
    This protocol serves as a guideline. Optimal reaction conditions must be individually determined.


    1. Thaw 10X Ammonium Buffer, dNTP mix, and primer solutions. It is important to mix the solutions completely before use to avoid localized concentrations of salts.


    2. Prepare a master mix according to Table 1. The master mix typically contains all the components needed for extension except the template DNA.
    The optimal MgCl2 concentration should be determined empirically but in most cases a concentration of 1.5 mM, as provided in the 1X Ammonium Buffer, will produce satisfactory results. Table 2 provides the volume of 25 mM MgCl2 to add to the master mix if a higher MgCl2 concentration is required.

    Table 1. Reaction components (master mix and template DNA)

    Component

    Vol./reaction

    Final Conc.

    10X Ammonium Buffer

    5 µL

    1X

    dNTP mix
    (12.5 mM of each)

    0.8 µL

    0.2 mM of
    each dNTP

    Primer A

    Variable

    0.1-0.5 µM

    Primer B

    Variable

    0.1-0.5 µM

    PR Polymerase

    1 µL

    2.5 units/reaction

    Distilled Water

    Variable

    - - - -

    Template DNA

    Variable

    0.1-0.5 µg/reaction

    Total volume

    50 µL

    - - - -

    Table 2. MgCl2 concentration in a 50µl reaction

    Final MgCl2 conc.
    in reaction (mM)

    1.5

    2.0

    2.5

    3.0

    3.5

    4.0

    4.5

    Additional volume
    of 25 mM MgCl2
    per reaction (µL):

    0

    1

    2

    3

    4

    5

    6

    3. Mix the master mix thoroughly and dispense appropriate volumes into reaction tubes. Mix gently, e.g., by pipetting the master mix up and down a few times.


    4. Add template DNA (0.1-0.5 mg/reaction) to the individual tubes containing the master mix.


    5. Program the thermal cycler according to the manufacturer's instructions. PR is a proofreading enzyme and requires an extension time of 1-2 min/kb. For maximum yield and specificity, temperatures and cycling times should be optimized for each new template target or primer pair.

    6. Place the tubes in the thermal cycler and start the reaction.
    Item#:
    ASPCRREAG5

    Pfu DNA Polymerase 
    Pfu DNA polymerase is a heat stable DNA polymerase which has 5' 3' DNA polymerase and 3' 5' exonuclease (proofreading) activities. Pfu DNA polymerase retains the high fidelity, sensitivity and processivity with an error rate six-fold lower than Taq DNA polymerase, and significantly lower than the error rates of most other proofreading enzymes or DNA polymerase mixtures (1). This product is supplied with the unique Intact Genomics 10x PCR reaction buffer, containing MgCl2, which produces a final Mg2+ concentration of 1.5 mM, and 5X Magic Enhancer that enables efficient amplification of GC rich templates up to 84%. The physical purity of this enzyme is 98% as assessed by SDS-PAGE with Coomassie® blue staining.

    Quality Control 
    Quality control is performed following the production of each new lot of product to ensure that it meets the quality standards and specifications designated for the product. Each lot is repeatedly compared side-by-side with leading competitors to ensure our products outperform the competitor before product launching. 

    Product Source
    E. coli strain expressing a Pfu DNA Polymerase gene from Pyrococcus furiosus.

    Contents & Storage

    1. Pfu DNA Polymerase
    2. 10x PCR Buffer with Mg²+
    3. 5x Magic Enhancer
    4. 10 mM dNTP (Cat. # 3312d, 3314d only)

    Store all contents at -20 °C.

    Storage Buffer 
    50 mM Tris-HCl, 50 mM KCl, 1 mM DTT, 0.1 mM EDTA, 50% Glycerol, pH 7.5 @ 25 °C.

    10X PCR Buffer with Mg2+ 
    100 mM Tris-HCl pH 9.0, 15 mM MgCl, 100 mM KCl, 80 mM (NH4)2SO4, 0.5% Igepal CA 630

    Unit Definition 
    One unit is defined as the amount of enzyme that incorporates 10 nmoles of dNTP into acid-insoluble form in 30 minutes at 72 °C.

    Protocol
    1. Thaw 10x PCR Buffer, dNTP mix, primer solutions, 5x Magic Enhancer (if required) and mix thoroughly before use. 
    2. Prepare a reaction mix according to the following table: The reaction mix typically contains all the components needed for PCR except the template DNA.

     

    PCR reaction set up:

    Template DNA

    xµl (0.01-0.5µMg)

    10x PCR Buffer

    10.0µl

    dNTP (10 mM)

    2.0µl

    Forward Primer

    xµl (0.1- 0.5µMM)

    Reverse Primer

    xµl (0.1- 0.5µMM)

    5x Magic Enhancer (optional)

    20µl

    Pfu DNA Polymerase (5 U/µl)

    0.5µl

    H2O up to

    100.0µl

    3. Mix the reaction mixture thoroughly.
    4. Add template DNA to the individual PCR tubes containing the reaction mixture. 
    5. Program the thermal cycler according to the manufacturerâs instructions. A typical PCR cycling program is outlined in the following table.

    PCR cycling conditions:

    Steps

    Temp.

    Time

    Cycles 

    Initial denaturation

    95 °C

    3-5 min

    1

    Denaturation

    94 °C

    30-60 sec 

      

    25-35

    Annealing

    52-66 °C

    30-60 sec 

    Extension

    72-74 °C

    1-2 min 

    Final extension

    72-74 °C

    10 min 

    1

    Hold 

    4-12 °C

    â

    6. Place the PCR tubes in the thermal cycler and start the cycling program.
    Item#:
    ASPCRREAG17

    HS Taq Polymerase Master Mix BLUE is a ready to use 2 X master mix. Simply add primers, template and water to successfully carry out primer extensions. The master mix is available in 100, 500, 1000, and 2500 reaction sizes and is available with two different buffer options (HS Buffer 1 and HS Buffer 2)


    HS Taq Polymerase is a modified form of Bullseye Taq, activated by heat treatment. This results in higher specificity and greater yields when compared to standard Taq. The added benefit of the HS Master Mix Blue is that it can be loaded directly onto the gel as there is no need to use separate loading dyes for subsequent electrophoresis and visualization. This cuts down on the chance of contaminating the component stocks and leads to better reproducibility.

    Composition of 2 X HS Master Mix Blue with Buffer 1
    Tris-HCl, pH 8.5, (NH4)2SO4,3 mM, MgCl2, .2 % Tween 20, .4 mM dNTPs, .2 units/uL HS Taq Polymerase, Inert Blue Dye, Stabilizer

    Composition of 2 X HS Master Mix Blue with Buffer 2
    Tris-Hcl, pH 8.5, Balanced KCl/(NH4)2SO4,3 mM, MgCl2, .2 % Tween 20, .4 mM dNTPD, .2 units/uL HS Taq Polymerase, Inert Blue Dye, Stabilizer

    Item#:
    ASPCRREAG15

    Cas9 Nuclease 
    Clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated (Cas) systems provide bacteria and archaea with adaptive immunity against viruses and plasmids by using CRISPR RNAs (crRNAs) to guide the silencing of invading nucleic acids. The CRISPR system consists of a short non-coding guide RNA (sgRNA) made up of a target complementary CRISPR RNA (crRNA) and an auxiliary transactivating crRNA (tracrRNA). The sgRNA guides the Cas9 endonuclease to a specific genomic locus via base pairing between the crRNA sequence and the target sequence, and cleaves the DNA to create a double-strand break. The location of the break is within the target sequence 3 bases from the NGG PAM (Protospacer Adjacent Motif). The PAM sequence, NGG, must follow the targeted region on the opposite strand of the DNA with respect to the region complementary sgRNA sequence

    Intact Genomics Cas9 Nuclease is the purified recombinant Streptococcus pyogenes Cas9 enzyme containing a nuclear localization signal (NLS) at the C-terminal for targeting to the nucleus. This enzyme is designed to perform CRISPR/Cas9-mediated genome editing. The physical purity of this enzyme is â¥98% as assessed by SDS-PAGE with Coomassie® blue staining.

    Quality Control 
    Quality control is performed following the production of each new lot of product to ensure that it meets the quality standards and specifications designated for the product. Each lot is repeatedly compared side-by-side with leading competitors to ensure our products outperform the competitor before product launching. 

    Cas9 nuclease is free from detectable RNase, Endonuclease (nicking) and non-specific DNase activities.

    Product Source 
    E. coli BL21 (DE3) strain expressing a Cas9 gene from Streptococcus pyogenes with an N-terminal 6xHis tag and C-terminal SV40 nuclear localization signal (NLS).

    Contents & Storage

    1. Cas9 Nuclease
    2. 10x Cas9 Nuclease Reaction Buffer

    Store Cas9 Nuclease and Buffer at -20 °C

    Storage Buffer 
    50 mM Tris-HCl, 50 mM KCl, 1 mM DTT, 0.1 mM EDTA, 50% Glycerol, pH 7.5 @ 25 °C

    1x Cas9 Reaction Buffer 
    20 mM HEPES, 100 mM NaCl, 5 mM MgCl, 0.1 mM EDTA, pH 6.5 @ 25 °C

    Functional Testing
    Cas9 Nuclease functional testing was done by in vitro DNA cleavage assay with the following protocol which gives more than 95% digestion of the substrate DNA as determined by agarose gel electrophoresis.
    1) Set up 30 µl reaction in a microcentrifuge tube on ice with the following combinations.

    Target DNA

    x µl (100ng)

    sgRNA

    x µl (4000ng)

    10x Cas9 Reaction Buffer

    3.0 µl

    Cas9 Nuclease

    1.0 µl (160ng)

    Add H2O up to

    30.0 µl


      

    2) Gently mix the reaction mixture and centrifuge briefly.
    3) Incubate at 37 °C for 60 min. 
    4) Add 1 µl RNase (4 mg/ml) 
    5) Incubate at 37 °C for 20 min.
    6) Run 0.7 to1% agarose TBE gel.

    Item#:
    ASPCRREAG14

    Taq DNA Polymerase, 2.0 Mix

    2.0 Master Mix Kit (1.5mM MgCl2)

    Cat. No.

    Reactions

    Taq DNA Polymerase

    MgCl2

    BE140301

    100

    2.0x Master Mix

    1.5 mM

    BE140303

    500

    2.0x Master Mix

    1.5 mM

    BE140306

    2,500

    2.0x Master Mix

    1.5 mM

    Store at -20°CFor in vitro laboratory use only

    General Description

    Bullseye Taq DNA Polymerase Mix is a ready-to-use 2.0X reaction mix. Simply add primers, template, and water to successfully carry out primer extensions and other molecular biology applications.

    Bullseye Taq DNA Polymerase, the NH4+ buffer system, dNTPs, and magnesium chloride are present in Taq DNA Polymerase Mix. Each reaction requires 25 µL of the 2.0X reaction mix. Simply add primers, template and water to a total reaction volume of 50 µL.

    Taq DNA Polymerase Mix offers several advantages. Set up time is significantly reduced. The chance of contaminating component stocks is eliminated. Reduction of reagent handling steps leads to better reproducibility. Standard tests can be set up with the confidence that results will be consistent every time.

      Composition of 2.0X Taq Master Mix

    • 150 mM Tris-HCl pH 8.5, 40 mM (NH4)2S04, 3.0 mM MgCl2, 0.2% Tween 20
    • 0.4 mM dNTPs
    • 0.05 units/µL Bullseye Taq polymerase
    • Stabilizer

      Suggested Protocol using Taq Master Mix

    This protocol serves as a guideline for primer extensions. Optimal reaction conditions such as incubation times, temperatures, and amount of template DNA may vary and must be individually determined.

      Notes:

    • Set up reaction mixtures in an area separate from that used for DNA preparation or product analysis.
    • The table below shows the reaction set up for a final volume of 50µL. If desired, the reaction size may be scaled down. Use 10 µL of the 2.0X master mix in a final volume of 20µL.
    • Important: Spin Taq Master Mix vials briefly before use.

      1. Set up each reaction as follows:

    Component

    Vol./reaction

    Final Conc.

    Taq Master Mix

    25 µL

    1X

    Primer A

    Variable

    0.1.1.0µM

    Primer B

    Variable

    0.1.1.0µM

    Distilled Water

    Variable

    - - - -

    Template DNA

    Variable

    Variable

    TOTAL volume

    50 µL

    - - - -

    2. Mix gently by pipetting the solution up and down a few times.

    3. Program the thermal cycler according to the manufacturer's instructions. For maximum yield and specificity, temperatures and cycling times should be optimized for each new template target or primer pair.

    4. Place the tubes in the thermal cycler and start the reaction.

    Item#:
    ASPCRREAG3

    Accuris PCR Promo!
    Buy One PR1001-HFHS-200, Get One Free of Equal or Lesser Value
    Please Contact Us for Full List of Qualifying Items
    Hurry! Promo Ends August 31, 2020

    Accuris High Fidelity Hot Start Master Mix

    • Leaves an A-overhang for TA cloning
    • Ideal for difficult, high GC content sequences
    • 10x fidelity of native Taq
    • Ideal for long PCR, up to 10kb targets

    Accuris High Fidelity Hot Start Mix is a hot start 2x formulation which provides excellent sensitivity in low-copy number assays with 10x higher fidelity than Taq polymerase. The 2x master-mix contains proprietary enhancers, an antibody mediated hot start mechanism, and a proof-reading component for trouble-free PCR reaction assembly and performance.

    The pre-optimized Hot Start Master Mix is supplied in a single tube, reducing the number of pipetting steps while improving throughput and reproducibility. The highly efficient buffer formulation and hot-start blend provide the ideal conditions for high-performance PCR and inactivity at room temperature thereby eliminating non-specific amplification.

    Item#:
    ASACCURISHIFIDHS
    • R-Taq DNA Polymerase 5 units/µL

      Cat. No.

      Units

      10X Ammonium Buffer (MgCl2 15mM)

      MgCl2

      25 mM

      BE200303

      500

      1.5 mL

      1.5 mL

      BE200304

      1,000

      2x 1.5 mL

      2x 1.5 mL

      BE200306

      2,500

      4x 1.5 mL

      4x 1.5 mL

      Store at -20°C. For in-vitro laboratory use only

       

      General Description

      Bullseye R-Taq DNA Polymerase is a thermostable recombinant DNA polymerase, which exhibits very high activity in primer extension and other molecular biology applications. R-Taq contains a red dye which provides easy and quick identification of reactions to which enzyme was added and allows confirmation of complete mixing. The inert dye has no effect on downstream processes. R-Taq is added directly to the reaction mix and is used in the same manner as standard Taq DNA Polymerase.

      Bullseye R-Taq DNA Polymerase has both a 5'®3' DNA polymerase and a 5'®3' exonuclease activity. The enzyme lacks a 3'®5' exonuclease activity. R-Taq DNA Polymerase leaves an A-overhang, which makes the enzyme ideal for TA cloning.

      • High performance thermostable DNA polymerase
      • Red dye identifies tubes which contain enzyme and confirms complete mixing of reagents
      • Leaves an A-overhang

      Unit Definition

      One unit is defined as the amount that incorporates 10 nmoles of dNTPs into acid-precipitable form in 30 minutes at 72°C under standard assay conditions.

      Storage Buffer

      Enzyme is supplied in 20 mM Tris-HCl pH 8.3, 100 mM KCl, 0.1 mM EDTA, 1 mM DTT, inert dye, 0.5 % Tweenà 20, 0.5% NP40, 50% glycerol.

      Component

      Vol./reaction

      Final Conc.

      10X Ammonium Buffer

      5 µL

      1X

      dNTP mix (12.5 mM each)

      0.8 µL

      0.2 mM each dNTP

      Primer A

      Variable

      0.1-0.5 µM

      Primer B

      Variable

      0.1-0.5 µM

      R-Taq DNA Pol

      1 mL

      5 units/reaction

      Distilled Water

      Variable

      - - - -

      Template DNA

      Variable

      0.1-0.5 µg/reaction

      TOTAL volume

      50 µL

      - - - -

      Table 2. MgCl2 concentration

      3. Mix the master mix thoroughly and dispense appropriate volumes into reaction tubes. Mix gently (e.g., by pipetting) the master mix up and down a few times.

      4. Add template DNA (0.1-0.5 mg/reaction) to the individual tubes containing the master mix.

      5. Program the thermal cycler according to the manufacturer's instructions.

      For maximum yield and specificity, temperatures and cycling times should be optimized for each new template target or primer pair.

      6. Place the tubes in the thermal cycler and start the reaction.

      7. After primer extension, load 5-10 mL of a 50 mL reaction directly on an agarose gel for analysis.


      Tween 20 is a registered trademark of ICI Americas, Inc.


      Final MgCl2 conc.

      in reaction (mM)

      1.5

      2.0

      2.5

      3.0

      3.5

      4.0

      4.5

      Additional volume

      of 25 mM MgCl2

      per reaction (µL):

      0

      1

      2

      3

      4

      5

      6

      10X Ammonium Reaction Buffer

      Tris-HCl pH 8.5, (NH4)2S04, 15 mM MgCl2, 1% Tween 20

      Quality Control

      Endonuclease, exonuclease and priming activities are not detected after 3 hours incubation of 1 mg of pUC19 plasmid DNA and 0.5 µg EcoR I digested lambda phage DNA at 72°C in the presence of 40 units of R-Taq DNA Polymerase.

      Suggested Protocol using R-Taq Polymerase

      This protocol serves as a guideline. Optimal reaction conditions such as incubation times, temperatures, and amount of template DNA may vary and must be individually determined.

      1. Thaw 10X Ammonium Buffer, dNTP mix, and primer solutions. It is important to mix the solutions completely before use to avoid localized concentrations of salts.

      2. Prepare a master mix according to Table 1. The master mix typically contains all the components needed for extension except the template DNA.

      The optimal MgCl2 concentration should be determined empirically but, in most cases a concentration of 1.5 mM, as provided in the 1X Ammonium Buffer, produces satisfactory results. Table 2 provides the volume of 25mM MgCl2 to add to the master mix if a higher MgCl2 concentration is required.

      Table 1. Reaction components (master mix & template DNA)

    Item#:
    ASPCRREAG4
    View: