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KingFisher™ Systems Consumables Thermo Scientific™

Optimize performance using Thermo Scientific™ KingFisher™ Systems Consumables, specifically designed for use with the KingFisher System and ideal for magnetic particle processing because of their low binding affinity for biomolecules. The specially-designed, disposable tip combs and microplates, which are made of polypropylene, enable excellent recovery of magnetic beads.

Consumables for KingFisher™ mL Systems Thermo Scientific™

Optimize system performance with Consumables for Thermo Scientific™ KingFisher™ mL Systems. Designed specifically for use with KingFisher mL instruments, these consumables utilize disposable plastics made of polypropylene. The plastics, tip combs and tube strips are ideal for magnetic particle processing due to their low-binding affinity for biomolecules, and their special design enables excellent recovery of magnetic beads.

LightShift™ Poly (dI-dC) Thermo Scientific™

Thermo Scientific LightShift Poly(dI-dC) can be used as competitor for nonspecific DNA binding proteins.

More Product Data
Transfer EMSA gels using the Pierce G2 Fast Blotter

Related Products
LightShift™ Chemiluminescent EMSA Kit
LightShift™ EMSA Optimization and Control Kit

Endotoxin Binding Reagent Thermo Scientific™

Thermo Scientific GeneJET Endotoxin Binding Reagent is a component of the GeneJET Plasmid Maxiprep Kit (K0491/K0492) and may be purchased separately. Provided in 60 mL aliquots.

KingFisher™ Plastics for 96 deep-well format Thermo Scientific™

Optimize your performance using Thermo Scientific™ KingFisher Plastics designed specifically for use with the KingFisher Flex, Duo Prime, Presto and Apex instruments. KingFisher instruments utilize disposable plastic plates and tip combs made of polypropylene, which are ideal for magnetic particle processing due to their low binding affinity for biomolecules.  Both the yield and quality of the isolated protein or nucleic acid are significantly improved with special plates and tip combs designed for KingFisher systems.

GeneArt™ Plasmid Premium Prep (high copy plasmid), TE Thermo Scientific™

The GeneArt® Gene Synthesis service offers chemical synthesis, cloning, and sequence verification of virtually any desired genetic sequence.

Platinum™ SuperFi II DNA Polymerase Invitrogen™

Invitrogen Platinum SuperFi II DNA Polymerase is a proofreading DNA polymerase that combines superior fidelity with an innovative buffer, enabling universal primer annealing for the highest success in PCR. It is ideally suited for cloning, mutagenesis, and other applications benefiting from supreme sequence accuracy.

Features of Platinum SuperFi II DNA Polymerase include:
• Exceptional >300X Taq fidelity
• Universal primer annealing at 60°C
• Superior specificity, sensitivity, and yields
• Robust amplification of difficult-to-amplify targets (e.g., those with suboptimal purity, ˃65% GC content, long PCR requirement)

Platinum SuperFi II DNA Polymerase is an engineered enzyme high processivity and increased resistance to PCR inhibitors. It also enables fast-cycling protocols and amplification of long targets (up to 20 kb). Platinum hot-start technology is based on proprietary antibodies that inhibit enzyme activity until the initial PCR denaturation step, preventing non-specific amplification and primer degradation. This technology also enables reaction setup at room temperature and provides increased sensitivity and yield.

Due to the unique composition of the SuperFi II PCR buffer, the annealing temperature is 60°C for most primer pairs designed following the general design rules. Isostabilizing molecules in the buffer increase primer-template duplex stability during the annealing step and contribute to enhanced specificity without the need to optimize the annealing temperature for each primer pair. With Platinum SuperFi II DNA Polymerase, different PCR assays can be cycled together using the same protocol with a universal primer annealing temperature and an extension step selected for the longest fragment to be amplified.

Applications of Platinum SuperFi II DNA Polymerase:
• High-fidelity PCR
• Cloning and sub-cloning
• Site-directed mutagenesis
• Amplification of GC-rich templates
• Template generation for sequencing
• High-throughput PCR
• Amplification of samples with suboptimal purity
• Long PCR
• Fast PCR

For increased convenience, we offer Platinum SuperFi II Master Mix with Platinum SuperFi II DNA Polymerase provided in a ready-to-use mixture along with SuperFi II PCR buffer and dNTPs, thus reducing the number of pipetting steps during PCR reaction setup. Platinum SuperFi II Green PCR Master Mix is also available, which additionally contains a density reagent and two tracking dyes for direct loading of PCR products on gels, further streamlining the PCR workflow from setup to final analysis.

View more information about Platinum SuperFi II DNA Polymerase products ›

LightShift™ EMSA Optimization and Control Kit Thermo Scientific™

The Thermo Scientific LightShift EMSA Optimization and Control Kit is an extraordinarily robust and sensitive system for performing electrophoretic mobility shift assays (EMSA) to identify and characterize protein-DNA binding interactions. The kit includes reagents for setting up and customizing DNA binding reactions and a control set of DNA and protein extract to test the kit system.

Features of theLightShift EMSA Optimization and Control Kit:

• Excellent for detecting low-abundance proteins in nuclear extracts
• Sensitivity that surpasses radioactive and digoxigenin methods
• Compatible with previously established binding conditions for popular DNA-protein interactions
• Includes EBNA control system to help new users develop a working assay and understand the methods used to confirm binding interaction specificity

The principle for LightShift EMSA Detection is similar to a Western blot. Biotin end-labeled duplex DNA is incubated with a nuclear extract or purified factor and electrophoresed on a native gel. The DNA is then rapidly (30 minutes) transferred to a positive nylon membrane, UV crosslinked, probed with streptavidin-HRP conjugate and incubated with the substrate. The protocol from labeling to results can be accomplished in a single day.

The interaction of proteins with DNA is central to the control of many cellular processes including DNA replication, recombination and repair, transcription, and viral assembly. One technique that is central to studying gene regulation and determining protein:DNA interactions is the electrophoretic mobility shift assay (EMSA).

The EMSA technique is based on the observation that protein:DNA complexes migrate more slowly than free DNA molecules when subjected to non-denaturing polyacrylamide or agarose gel electrophoresis. Because the rate of DNA migration is shifted or retarded upon protein binding, the assay is also referred to as a gel shift or gel retardation assay. Until conception of the EMSA protein:DNA interactions were studied primarily by nitrocellulose filter-binding assays.

All that is needed to perform the assay is purified DNA target that has been end-labeled with biotin, the protein extract to be tested, nylon membrane and basic electrophoresis equipment. DNA targets can be synthesized with 5' or 3' biotin labels or they can be labeled after synthesis using the Thermo Scientific Biotin 3' End DNA Labeling Kit (Product No. 89818). Nuclear, cytosolic or whole cell protein extracts can be obtained by a variety of methods, including the Thermo Scientific NE-PER Nuclear and Cytoplasmic Extraction Reagent Kit (Product No. 78833).

More Product Data
Transfer EMSA gels using the Pierce G2 Fast Blotter

Related Products
LightShift™ Chemiluminescent EMSA Kit
LightShift™ Poly (dI-dC)

Quant-iT™ dsDNA Assay Kit, broad range Invitrogen™

The Quant-iT™ Broad-Range dsDNA Assay Kit makes DNA quantitation easy and accurate. The kit provides concentrated assay reagent, dilution buffer, and pre-diluted DNA standards. The assay is highly selective for double-stranded DNA over RNA, and in the range of 2-1000 ng of DNA, the fluorescence signal is linear. A Quant-iT™ dsDNA Assay Kit with a range of 0.2-100 ng of dsDNA is available (Q33120), and we also offer a Quant-iT™ RNA Assay Kit (Q33140).

Check out our products for nucleic acid purification

MagMAX™ FFPE DNA/RNA Ultra Kit Applied Biosystems™

The MagMAX™ FFPE DNA/RNA Ultra Kit is designed for sequential isolation of DNA and RNA from the same formaldehyde- or paraformaldehyde-fixed paraffin-embedded (FFPE) tissue sample. The DNA and RNA are recovered in separate eluates, and both are compatible with a broad range of applications, including real-time PCR and next-generation sequencing. The isolation of RNA and DNA from the same FFPE sample makes this kit an ideal method of sample preparation for Oncomine™ Comprehensive and Oncomine™ Focus assays.

Features of the MagMAX FFPE DNA/RNA Ultra Kit include:
• Flexible design that permits both manual and automated isolation of RNA and DNA
• Minimal requirement of 5 µm section or curl input
• Capable of processing 40 µm FFPE sections or curls; greater than 40 µm sections can be processed with an alternative protocol
• Compatible with targeted RNA and DNA sequencing panels
• Contains Dynabeads™ MyOne™ Silane for consistent isolation of RNA, microRNA, and DNA
• Alternative protocols for the isolation of RNA or DNA only

Unlocking the information contained in FFPE samples is essential to cancer research. Common nucleic acid isolation methods require you to choose which nucleic acid to recover from a given sample, with loss of the other nucleic acid (and any information that may have resulted from its analysis). Alternatively, the DNA and RNA are co-purified, or the sample is split in half so that each can be purified.

More information from less sample input
The MagMAX FFPE DNA/RNA Ultra Kit protocol addresses these issues by first isolating DNA from the FFPE sample and then recovering the RNA from the DNA-depleted supernatant. The nucleic acids are prepared as separate eluates and are then ready for downstream analysis. This allows for a fuller analysis of precious samples, including investigation of important biomarkers, such as hotspot mutations, CNVs, gene fusions, and indels.

Benefits of MagMAX magnetic bead–based purification
The magnetic bead–based purification format of the kit allows for easy scaling of the number of samples processed either manually with a magnetic stand or with a magnetic particle processor such as the KingFisher™ Duo Prime or KingFisher™ Flex processor.

Magnetic beads offer many benefits compared to other technologies for isolating nucleic acids. Beads bind the RNA and DNA more efficiently than glass fiber filters, resulting in higher and more consistent yields. Additionally, because filters and vacuum manifolds are not used, there is no risk of undigested tissue particulates clogging these items during the extraction process. This clogging issue is of particular concern with fibrous FFPE tissue samples that that are not fully digested during the proteinase K treatment.

Optimized bead surface chemistry
The Dynabeads MyOne Silane utilized in the MagMAX FFPE DNA/RNA Ultra Kit have an optimized silica-like surface chemistry with high specific surface area that allows efficient kinetics and high sensitivity in nucleic acid capture. These aspects make Dynabeads MyOne Silane ideal for the recovery of both RNA and DNA targets found in FFPE samples. Once captured, both of these nucleic acids are then eluted separately in volumes of 50 µL and are ready for downstream applications.

Neutralization Solution for MagJET Plasmid DNA Kit Thermo Scientific™

Thermo Scientific MagJet Neutralization Solution is a component of the MagJET Plasmid DNA Kits (K2791/2792) and may be purchased separately. Provided in 25 mL aliquots.

KingFisher™ Plastics for 96 standard and PCR formats Thermo Scientific™

Optimize your performance using Thermo Scientific™ KingFisher Plastics designed specifically for use with the KingFisher Flex, Duo Prime, Presto and Apex instruments. KingFisher instruments utilize disposable plastic plates and tip combs made of polypropylene, which are ideal for magnetic particle processing due to their low binding affinity for biomolecules. Both the yield and quality of the isolated protein or nucleic acid are significantly improved with special plates and tip combs designed for KingFisher systems.

pcDNA™3.1/nV5-DEST Mammalian Expression Vector Invitrogen™

To fit all of your expression needs, Invitrogen offers state-of-the-art Gateway® destination vectors for expression in E. coli, insect, yeast, or mammalian cells, as well as for production of native protein or N- or C-terminal fusion proteins. All Gateway® destination vectors have attR sites for recombination with any attL-flanked fragment, regardless of whether it is an entry clone or an Ultimate™ RF Clone. The following table lists the wide range of destination vectors available.

Additional materials required, available separately: Gateway® entry clone, Gateway® LR Clonase® enzyme mix, and reaction buffer.

pPICZ A, B, & C Pichia Vectors Invitrogen™

The pPICZ A, B, & C Pichia Vectors are designed for simple cloning and selection, high-level expression, and rapid detection and purification of the recombinant protein. These vectors contain the Zeocin™ resistance gene for direct selection of multi-copy integrant strains. By selecting with increasing amounts of Zeocin™, strains with multiple copies of your gene of interest integrated into the genome are obtained. Increasing the number of copies of the gene of interest in a recombinant Pichia strain can result in higher expression levels. The vectors are included in the EasySelect™ Pichia Expression Kit (Cat. No. K1740-01).

Features of the pPICZ vectors include:

• Inducible AOX1 promoter for high-level expression in Pichia pastoris
c-
myc epitope tag for convenient detection with an Anti-myc Antibody
• C-terminal polyhistidine (6xHis) tag for rapid purification with nickel-chelating resin and detection with an Anti-His(C-term) Antibody
• Zeocin™ resistance for direct selection of multi-copy integrants

Wash Solution I (concentrated) Thermo Scientific™

Thermo Scientific GeneJET Wash Solution I (concentrated) is a component of the GeneJET Plasmid Maxiprep Kit (K0491/K0492) and may be purchased separately. Provided in 360 mL aliquots.
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