Gene Expression Analysis
One World Biotech provides a focused set gene expression and genotyping assays for detection and accurate quantification of nucleic acids.
Reverse Transcription Quantitative PCR (RT-qPCR) is often considered to be the gold standard for measuring differences in gene transcripts, however data quality and reliability can be affected by variability in the RNA template, assay design and data normalization.
We understand that high-quality qPCR data can be generated by adhering to the following four guidelines: use of RNA of highest integrity, accurate determination of RNA concentration, use of a suitable reference gene for normalization, and precision pipetting. We follow MIQE guidelines and supply complete information about each experiment.
Our RNA services include:
> Purification of total RNA, mRNA, and microRNA
> Extraction of RNA from Paraffin-Embedded (FFPE) Tissue
> TaqMan and TaqMan Low Density Array (TLDA) Assays
> In Vitro Transcription and mRNA Synthesis
> RT-PCR and 5’ and 3’ RACE-PCR
> Ribonuclease Protection Assay (RPA)
> RNA Electrophoresis Mobility Shift Assay (EMSA)
Our DNA services include:
> Methylation Sensitive-High Resolution Melting (MS-HRM)
> Methylation Analysis by Methyl PCR Array
> Genotyping by Real-Time PCR
> Single Nucleotide Polymorphism (SNP)
> Transgene Insertion Site and Transgene Copy Number
> DNA Gel Shift Assay
Gene Cloning
We create custom recombinant DNA plasmid and viral constructs by combining various sequence elements including: constitutive and inducible promoters, natural, synthetic, reporter, hybrid and mutant genes, 5' and 3' untranslated sequences, terminators, post-transcriptional regulatory elements, matrix attachment regions and other sequences to study gene function and to develop cell lines or organisms that produce biologically active protein products.
We can meet all of your custom gene cloning, subcloning, mutagenesis, and plasmid DNA purification needs. We have constructed hundreds of plasmid and viral vectors including:
> Reporter Gene Plasmids with various GFPs
> Promoter and Subcellular Localization Signal Constructs
> Cre-Lox and Cre-FLEX Constructs
> Tetracycline-Inducible Expression Plasmids
> Gene Targeting Constructs
> siRNA and microRNA Constructs
> Knock-in, Knock-out and Knock-down Plasmids
> Viral Vector Constructs (AAV and Lentivirus)
Cloned genes are often introduced into mammalian cells to develop cell-based assays. One World Biotech transforms and characterizes cell lines by western, ELISA, FACS, RT-PCR, and qPCR. We also develop and validate cell-based assays for a variety of applications including cytotoxicity, siRNA, reporter gene expression, and receptor binding.
> Transient Transfection
> Stable Transfection
> Single Cell Clonal Selection