NucleoWay™ Molecular Cloning

OneWorld Biotech, Inc. is now pleased to offer expanded and cost effective NucleoWayTM Molecular Cloning services to the wider scientific community. We have been helping biotech and pharma clients with custom gene cloning and vector construction, RNA and mRNA synthesis, plasmid DNA and RNA purification, and other DNA/RNA analysis projects over the past nine years.

OneWorld Biotech Advantage

  • We have proven expertise in delivering highest quality DNA and RNA services: constructed thousands of plasmid and viral vectors and produced multi milligram quantities of RNA and mRNA for transfection, structural and biochemical studies, and controls and standards.
  • NucleoWayTM Molecular Cloning services provide solutions for your DNA and RNA project needs. OneWorld Biotech will design and produce complex DNA constructs for nearly any purpose.
  • We use best-in-class technology platforms to provide most effective solutions for your research needs.

DNA Services

Custom Gene Cloning and Vector Construction

  • Cloning of genes and synthetic genes from any species in the vector of your choice — in our in-house vector or in a new vector created just for your project.
  • Experienced scientists at OneWorld Biotech have cloned genes and constructed hundreds of protein expression vectors for bacterial, yeast, insect, and mammalian cell expression.
  • Use our expertise to create plasmids for your research in a fast and cost- effective manner and focus on your core capabilities.
  • We supply sequence confirmed plasmid DNA, plasmid map and Applied Biosystems Sequencer chromatogram files.

Cloning by Restriction Enzyme Digestion: Digest plasmid containing the insert, gel isolate insert, ligate to vector at same sites or at blunt ends.
Cloning by PCR Amplification: PCR amplify insert, digest with restriction enzymes, and ligate to vector at same sites or clone by TA or blunt end cloning.
Cloning by Gene Synthesis: Clone synthetic gene or sequence using restriction enzyme sites or by TA or blunt end cloning.
Sequence and Ligase Independent Cloning: Amplify gene using special PCR primers and anneal insert and linear vector.
Cloning by Gibson Assembly: Use exonuclease-based method to seamlessly assemble several DNA fragments in correct order for gene synthesis, synthetic biology, and novel expression vectors.
Cloning by RT-PCR: Isolate RNA/mRNA from cells and tissues, reverse transcribe RNA using gene specific primers and clone into any plasmid.
Protein Expression Vectors: Vectors created by cloning genes, marker genes, chimeric genes, fusion genes, splice variants, mutants, epitope and purification tags, promoters, chimeric promoters, terminators, antibiotic resistance genes, insulators, and MARS to create vectors for transient and stable expression in E. coli, yeast, insect, and mammalian cells.
Viral Expression Vectors: AAV, Adeno, Lenti, Baculovirus, and other viral vectors with combination of genes, promoters and tags.
Gene Targeting Vectors: Vectors for homologous recombination, knock-in, knock-out, Cre, FLP, and Flox constructs for genome integration.
Inducible Expression Vectors: Conditional and inducible systems for a temporal and spatial controlled activation of genes including Tet-OnTM, Tet-OffTM, GeneSwitch±, Ecdysone-inducible, and tamoxifen-inducible.
Small RNA Vectors: Clone siRNA, miRNA and shRNA in plasmids with pol II or pol III promoters for transient and stable expression.

Plasmid DNA Purification and other services

  • Fast and cost effective high purity plasmid DNAs for research and preclinical applications.
  • Transfection grade and Endotoxin-Free grade plasmid DNAs from micrograms to milligrams scale for various applications.
  • Industry standard anion-exchange chromatography for purification and additional polishing step when needed.
  • Quality control specifications includes: A260/A280 ratio; >1.8, Homogeneity; predominantly supercoiled, Genomic DNA; not detected on agarose gel, RNA; not detected on agarose gel, Restriction enzyme digestion; matching to expected fragments, and Endotoxin-free; as specified.

We are a truly custom services company and our services can be tailored to meet your specific requirements. We can fulfill your needs of recombinant DNA constructs and other molecular biology experiments including DNA- and RNA-based assays and analyses. To learn more about how we can accelerate your research and to obtain a no-obligation quote, contact us today.

  • DNA- and RNA-based assays and analyses.
  • Genomic DNA purification from any type of cells and tissues.
  • DNA, RNA and Protein isolation from same sample.
  • Purification of PCR or restriction size fragments from agarose gels.
  • Minipreps, endotoxin-free minipreps, and 96-well plate preps.
  • Mutagenesis: Additions, deletions, substitutions, alanine scanning, site-specific and random mutagenesis.

RNA Services

RNA Services

We offer a comprehensive set of RNA services and have the flexibility to create specialized methods for unique scientific objectives.

  • Synthesis of milligram quantities of RNA and mRNA for transfection, structural and biochemical studies, and controls and standards.
  • Highly pure mRNA for studies on induced pluripotent stem cells, T-cell therapy, vaccines, microinjection, and in vivo therapeutic protein expression.
  • RNA extraction from any type of cells, tissues and paraffin-embedded samples and analysis by RT-PCR, Northern, nuclease mapping, and RNA-protein interaction.
  • RNA quality control by Agilent 2100, NanoDrop 2000, HPLC, and denaturing gel electrophoresis.

Our RNA services are described below in general terms. Working with OneWorld Biotech, you will experience first-hand our commitment to exceptional service.

RNA and mRNA Synthesis, and Quality Control

OneWorld Biotech’s comprehensive synthesis services allow us to use the latest technologies to provide clients with the highest quality RNA using T7, T3, SP6 RNA polymerases, RNA with homogenous 5’ and 3’ ends, RNA produced in E. coli, and mRNA with 5’ cap and 3’ poly (A) tail. RNAs of 20 to 5000 nucleotides with synthesis scale of 100 ug to 100 mg are available. Projects are tailored to meet client specifications.

Our goal is to exceed our client’s expectations with respect to the quality of work. We employ the Quality by Design approach at all steps of synthesis and purification. Quantity (NanoDrop 2000, Fluorometric), purity (NanoDrop 2000, denaturing gel electrophoresis, HPLC), and integrity (Agilent Bioanalyzer, denaturing gel electrophoresis) are evaluated and QC data is shared with our clients.

RNA Isolation and Analysis

The success of all of gene expression analysis methods from Northern blotting to RT-PCR and RNAseq is dependent on the quality of RNA isolated. We can assist with your gene expression analysis research by providing high quality RNA as well as full-length cDNA prepared for optimum performance in analysis. RNA, poly(A) RNA, and microRNA isolation from all types of cells and tissues, including demanding samples such as formalin-fixed and paraffin-embedded tissues (FFPE) are prepared to the highest standards, and to whatever scale required.

RNA extracted from cells and tissues using industry standard reagents and in-house expertise has been used for gene expression analysis, cDNA library construction and screening in HEK293 cells to identify a drug target, and cloning of low abundance RNAs by reverse transcription.