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Enzymes for Molecular Biology

Reaction Cleanup and Improving Yields

Choose the right enzyme for cleaner samples and faster processing

Reagents and reactants that are left behind can hinder downstream processes. Use the right enzyme preparation to clean up your samples before taking the next step. Speed up and enhance amplification and transcription with some not-so-secret additives. 

Enzymatic cleanup of DNA samples with Exonuclease I, DNase I, RNase A, Proteinase K and other nucleases and proteases removes residual primers, nucleotides and enzymes prior to SNP analysis, next-generation sequencing, Sanger DNA sequencing, bioprocessing or other downstream procedures.

Proteinase K is a broad-spectrum endopeptidase widely used for the digestion of proteins, including DNases and RNases. A digestion step with the enzyme is routinely applied during nucleic acid preparation without affecting the integrity of isolated DNA or RNA. Proteinase K is active under a wide range of reaction conditions, including elevated temperatures and the presence of SDS.

Proteinase K NGS Grade is available for the most demanding applications, including preparation of nucleic acids for next-generation sequencing. Extensive purification yields a high-quality enzyme product with increased specific activity, significantly increased solubility (2.5 fold) and remarkable purity with DNA content ≤0.1 pg/mg. Proteinase K NGS Grade is free of exonucleases, endonucleases and ribonucleases.

The TAGZyme DAPase enzyme and TAGZyme system is used for His-tag removal from proteins containing an intrinsic DAPase stop point (expressed using the TAGZyme pQE-2 vector) or from proteins that contain an engineered glutamine stop point.

A comprehensive range of housekeeping enzymes is available for cleavage and cleanup of residual reagents.

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May it be for your reaction setup, converting DNA & RNA mass and moles or calculating volumes of solutions, we have tools made for you.

Improved PCR yields and quality of templates may be achieved with the use of proteins that counteract PCR inhibitors and by adding specialized DNA-binding proteins to amplification and sequencing reactions. The DNA-binding protein, gene 32 protein from bacteriophage T4 (T4gp32), increases PCR amplification efficiency with a number of diverse templates. In addition, using E. coli single-stranded DNA-binding protein (SSB) in DNA sequencing reactions increases the resolution of sequencing runs.

An increased rate of in vitro transcription is made possible by treatment with inorganic pyrophosphatase, an essential component of reactions for RNA preparation. This enzyme cleaves pyrophosphate into two phosphate molecules and prevents pyrophosphate from precipitating with magnesium.

* This highly exergonic reaction can be coupled to unfavorable biochemical transformations to drive these transformations to completion.

†Pushes chemical equilibrium toward the synthesis of DNA by removing pyrophosphate from the reaction. This enzyme cannot be heat-inactivated, and retains full activity after incubation at 100°C for hours.

UDG-mediated strand cleavage is an important tool in molecular biotechnology, allowing for controlled and location-specific cleavage of single- and double-stranded DNA chemically or enzymatically synthesized with single or multiple incorporations of deoxyuridine. 

UDG treatments can reduce sequence artifacts, eliminate PCR carry over and generate specific gaps in DNA.

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FAQs about reducing artifacts and reaction cleanup

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