Bullseye EasyScript™ Reverse Transcriptase

Application

• Synthesis cDNA froma single-stranded RNA or DNA primer extension
• Sequencing dsDNA
• cDNA library
• Template production for use in PCR
• 3'-end labeling of duplex DNA via end-filling reactions

EasyScript™ Reverse Transcriptase is a genetically modified form of Moloney Murine Leukemia Virus Reverse Transcriptase (M-MLV). Both the EasyScript™ and EasyScript™ Plus are RNase H deficient (negative). The enzyme is purified from bacteria h o s t as a single holoenzyme of 71 kDa with the capacity of first-strand cDNA synthesis of template up to 9kb and can be used for routine cDNA synthesis.

Kit Components

Storage Buffer
 
50 mM Tris-HCl (pH 8.3), 100 mM NaCl, 0.1 mM EDTA, 5 mM DTT, 0.1% (v/v) Triton X-100, and 50% (v/v) glycerol.

Storage
 
Store at -20°C in a frost-free freezer. Multiple freezing and thawing of RNA should be avoided. Keep RNA on ice all the time. It is recommended that the first strand cDNA synthesis is carried out under conditions where RNase contamination has been eliminated.

General Protocol

RT-PCR reactions should be assembled in a RNA-free environment. The use of "clean", automatic pipettes designated for PCR and aerosol resistant barrier tips are recommended.

1. Thaw template RNA and all reagents on ice. Mix each solution by vortexing, and centrifuge briefly to collect residual liquid from the sides of the tubes. 
2. Prepare the following reaction mixture in a PCR tube on ice:

3. Incubate at 25°C for 10 minutes if random primer is used. Omit this step if Oligo(dT) primer or sequence specific primer are used.
4. Incubate the mixture at 42°C for 60 minutes.
5. Stop the reaction by heating at 85°C for 5 minutes. 
6. Chill on ice. The newly synthesized first-strand cDNA now can be used directly for PCR amplification.

Notes:
1. Isolation of poly(A)+ RNA from total RNA is not mandatory; however, doing so may improve the yield and purity of the final product. 
2. RNA sample must be free of contaminating genomic DNA.
3. Unlike the oligo(dT) priming, which usually requires no optimization, the ratio of a randomprimer to RNA is critical in terms of the average length of cDNA synthesized in the reaction. Increasing the ratio of random primer/RNA will result in higher yield of shorter (~500 bp) cDNA, whereas decreasing this ratio will produce longer products.

4. The synthesized cDNA should be stored at -20°C.

Bullseye 1Kb DNA Ladder, Logic

• Ready to use
• Contains 16 DNA bands: 100bp-10Kb
• 920ng DNA/ 6 uL/loading
• Easy quantification of DNA fragments
• Stable at room temperature
• Supplied with 6x sample loading buffer

Bullseye 1Kb DNA Ladder, Logic contains 16 DNA bands ranging from 100bp to 10Kb, formulated so that each band contains an amount of DNA that correlates logically to its size, allowing the user to estimate both the size and the quantity of specific fragments at a glance. It is particularly useful for protocols such as probe labeling, DNA sequencing and optimizing insert/vector ratio in ligation reactions, where DNA concentration must be taken into consideration.

Size: 1200 uL
Storage: Store at -20°C.
Concentration: 920 ng/6 uL

Loading Buffer Composition:

• 10mM Tris-HCl
• 1mM EDTA (pH 8.0)
• 0.02% Bromophenol blue
• 0.02% Xylene cyanol
• 5% Glycerol

Usage: Add 6 uL of Bullseye 1Kb DNA Ladder, Logic directly to wells designated for markers, which will yield the amount as indicated in the picture. You may use more than 6 uL of ladder, depending on well size and level of dye used to visualize the bands. Calculate the amount for each band according to your loading volume.

 Bullseye Individual dNTP's

• Ready-to-use molecular grade dNTP solution for use in DNA polymerization, DNA labeling and sequencing processes
• High purity: >98% by HPLC
• Supplied in solution at pH 7.5
• dNTPs are stable at -20°C, avoid multiple freeze/thawing (For long-term usage, aliquoting is recommended)
• Functionally tested with thermostable polymerases

• Mix of dATP, dCTP, dGTP, dTTP
• Each nucleotide is at a concentration of 100 mM
• Ready-to-use molecular grade dNTP solution for use in DNA polymerization, DNA labelling and sequencing processes
• High purity: > 98% by HPLC
• Supplied in solution at pH 7.5
• dNTPs are stable at - 20°C, avoid multiple freeze/thawing (For long-term usage, aliquoting is recommended)
• Functionally tested with thermostable polymerases 

dNTP SET

(dATP, dCTP, dGTP, dTTP)

100 mM

This brand is being discontinued and will only be available while supplies last.
 

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Bullseye DNA Safe Stain Plus

• Higher sensitivity
• Use in the same way as EtBr in agarose gel electrophoresis
• Detection of DNA and RNA
• Safest DNA stain by far
• Low cost
• 10,000X
• Excitation wavelengths at 290nm and 490nm

DNA SafeStain Plus is a highly sensitive green fluorescent DNA/RNA staining reagent for detecting nucleic acids in agarose and polyacrylamide gels. This unique stain gives high sensitivity for detection of double-stranded or single-stranded DNA and RNA. Gels can be post-stained or the stain can be added to gels during gel casting or to the gel running buffer. DNA SafeStain Plus has two excitation wavelength peaks at about 290nm and 490nm, and an emission wavelength at 530nm, making it compatible with a standard UV light box, a blue-light transilluminator, or a gel reader equipped with visible light excitation; such as, a 488 nm laser-based gel scanner.

DNA SafeStain Plus is in a 10,000X concentrated format that can be easily diluted 10,000 times for use in precast gel staining, or 5,000 times for use in post gel staining.

Pfu 2x Master Mix 
Pfu DNA Polymerase 2x master mix is ready to use premix which contains Pfu DNA Polymerase, dNTPs, MgCl2 and stabilizers with optimized reaction buffer. It has been optimized for routine PCR applications. 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. Pfu 2x Master Mix product is supplied with the unique Intact Genomics 5x Magic Enhancer that enables efficient amplification of GC rich templates up to 84%.

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 2x master mix
2. 5x Magic Enhancer

Store all contents at -20 °C.

1x Master Mix Composition 
10 mM Tris-HCl pH 9.0, 50 mM KCl, 1.5 mM MgCl, 0.2 mM dNTPs, 5% Glycerol, 0.08% Igepal CA 630, 0.05% Tween-20, 100 Units/ml Pfu DNA Polymerase.

Protocol
1. Prepare a reaction mix according to the following table:
  

2. Mix the reaction mixture thoroughly.
3. Program the thermal cycler according to the manufacturer's instructions.
4. A typical PCR cycling program is outlined in the following table.

5. Place the PCR tubes in the thermal cycler and start the cycling program.

6. Analyze 5 µl of PCR products by agarose gel electrophoresis. 

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.

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.

This product is no longer available, please contact us for other options.
 

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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.

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.

Bullseye PREMIUM HS-Taq DNA Polymerase Master Mix Blue

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

Bullseye Taq DNA Polymerase

Bullseye Taq DNA Polymerase is the most popular thermostable enzyme used in DNA amplification experiments. This high performance Taq DNA polymerase is specifically purified to produce excellent yields with little or no background. Its outstanding activity and unique thermal DNA amplification properties make it one of the best-value DNA polymerases available.

• Robust performance
• Leaves 3' A overhang
• Stable at all storage temperatures
• Ideal for general PCR, genomic analysis and TA cloning

Storage: -20°C

Quality Control
Taq DNA Polymerase is highly purified, free of contaminating endonucleases, exonucleases and nicking activity. For endonuclease assay, 1 ug of Lamda/Hind III DNA is incubated with 20 units of enzyme in assay buffer at 75oC for 16 hours with no visible contaminating activity observed. Also, every lot is tested for its performance consistency.

Unit Definition
One unit incorporates 10n moles of 4 radioactive labeled dNTPs into acid-insoluble material in 30 minutes at 74°C.

Storage Buffer
5 units / uL in 50 mM Tris-HCl (pH 8.0), 100 mM NaCl, 0.1 mM EDTA, 1 mM DTT, 50% glycerol, 0.5% TritonX-100, and 0.5% NP-40.
10x Reaction Buffer 100 mM KCl, 100 mM Tris HCl (pH9.0), 80 mM (NH4)2SO4, and 1.0% Triton X-100. (Mg++free): is optimized for use with 200 uM dNTPs.

Magnesium Chloride
25 mM MgCl2: In general, 1.5 mM MgCl2 is recommended; this may vary with different conditions and primer sets.
 
*For laboratory research only. Not for clinical applications. 

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This brand is being discontinued, please contact us for other options.
 

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Bullseye PREMIUM GC Rich Optimized DNA Polymerases and Kits

Key Features

• For amplification of DNA targets with high GC content
  
• Convenient reaction set-up at room temperature
  
• High specificity, sensitivity and product yield
  
• Detection of low abundance targets
  
• Diminished formation of non-specific product

  Composition of GC HS 2x Master Mix I
  

• HS DNA Polymerase
  
• Optimized buffer components, 3.0 mM MgCl
  
• dNTPs
  
• Enhancer