2x Master Mix Kit (1.5 mM MgCl2) 

Bullseye Taq DNA Polymerase Mix is a ready-to-use 2x reaction mix. Simply add primers, template, and water to successfully carry out primer extensions and other molecular biology applications.

Bullseye Taq polymerase, the NH4+ buffer system, dNTPs, and magnesium chloride are conveniently present in the Taq DNA Polymerase Mix. (Inert Red Dye is present in BE180303 only)

Bullseye 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 2x Taq Master Mix

150 mM Tris-HCl pH 8.5, 40 mM (NH4)2S04, 3 mM MgCl2, 0.2% Tween 20Ò

4 mM dNTPs

2 units/µL AS ONE Taq polymerase

Inert Red Dye & Stabilizer (BE180303 only)

Bullseye RT 2X Master Mix

• Up to 9kb cDNA synthesis
• Ensures sample to sample consistency
• Large RNA sample volume capacity
• Ready to use

Size: 100 rxns

RT 2X Master Mix is a proprietary, ready-to-use master mix for first-strand cDNA synthesis in a 2X concentration. This optimized reaction mix contains ribonuclease inhibitor, dNTPs, and a balanced concentration for oligo(dT) and random primers. The ribonuclease inhibitor effectively protects RNA template from degradation. The oligo(dT) anneals selectively to the poly(A) tail of mRNAs and the random primers do not require the presence of poly(A) and they are utilized for the transcription of mRNA 5-end regions. The resultant cDNA can be directly used as template in different PCR experiments.

Kit Components

Storage

• Store at -20°C in a frost-free freezer.

Application

• cDNA synthesis
• Construction of cDNA libraries
• Generation of probes for hybridization 

Protocol

1. Thaw RNA templates and all reagents on ice. Mix each solution by vortexing.
2. Assemble the following components in a tube on ice, and mix well:

1. Heat the mixture at 65°C for 5 mins and incubate on ice for at least 1 min.
2. Collect all components by a brief centrifugation and add 1 µl of the EasyScript RTase to the tube.
3. Mix well and collect all the components by a brief centrifugation.
4. Incubate the tube at room temperature for 10 min for annealing.
5. Perform cDNA synthesis by incubating the tube for 50 min at 42°C.
6. Stop the reaction by heating it at 85°C for 5 min.
7. Chill on ice. The newly synthesized first-strand cDNA is ready for immediate downstream applications. 

Bullseye EasyScript cDNA Synthesis Kit

Application

• First strand cDNA synthesis for PCR
• Construction of cDNA libraries
• Generation of probes for hybridization

EasyScript cDNA Synthesis Kit is a complete system for the efficient synthesis of first strand cDNA from RNA templates. The recombinant RNasin Ribonuclease Inhibitor, supplied with the kit effectively protects RNA template from degradation.

The kit is also supplied with both oligo(dT) and random primers. The oligo(dT) anneals selectively on the poly(A) tail of mRNA. Random primers do not require the presence of poly(A). Therefore, they can be used for transcription of the 5'-end regions of mRNA. Gene-specific primers may also be used with the kit. The first strand of cDNA can be directly used as a template in PCR.

EasyScript Reverse Transcriptase (RTase) within the kit 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 host 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 random primer 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 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.

 PCR Fast Plate, 96x0.1ml

• Optimized for fast PCR and qPCR compatible
• Ultra-thin and consistent wall thickness for precise thermal transfer
• Made from 99.9% virgin polypropylene
• 240ul well capacity (working capacity dependent on closure method)
• Designed for ABI thermal cyclers 7500 and 7900, 9800, Veriti, and StepOne
• Certified DNAse/RNAse and pyrogen-safe