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PCR / RT PCR Services

PCR Cloning and shRNA-encoding vector construction 
We provide cloning services, DNA sequencing, alignment, and plasmid construction. Customers are welcome to send us the Gene ID/Genbank/Swiss-Prot number, the gene/protein sequence, or a plasmid containing a gene of interest.

shRNA Design and Synthesis
We offer a complete service for preparing DNA templates and synthesizing hairpin shRNAs. Purification of the resulting shRNAs and preparation for transfection into cultured cells are offered as well. mRNA Amplification uses the T7 promoter (tagged polydT) primer for priming first strand cDNA synthesis. Double-stranded cDNA is used as a template for in vitro transcription.

In addition to standard ribonucleotides, company scientists also incorporate modified nucleotides (e.g., aminoallyl-UTP) to produce labeled RNA probes and amplifies the original mRNA more than 1 million-fold. RNA is also purified and prepared for all additional applications.

Gene Knockdown by shRNAs

The customer provides the name and accession numbers of the target genes, mails the cells and primary antibodies for Western blotting analysis to us, and the lab team does the rest . Our service includes design, construction, and confirmation of the 3′-5′ sequence the shRNA vectors for every target gene. Design and synthesis of the SYBR-labeled qPCR primers for quantitation of target gene expression, shRNA transfection into the cell line supplied by the customer, and targeted gene expression analysis by qPCR at 24, 48, and 72 hr post-transfection and Western blot at 48 and 72 hr after transfection are performed by company scientists. Transfection controls includes the negative vector control with scrambled sequences (21 bp) and a GFP protein vector control for assessment of transfection efficiency.

Real-Time PCR Services (RT PCR)

Quantitative PCR (QPCR), is a DNA sequence detection and quantitation method extensively used for DNA copy number detection, gene expression, SNP genotyping, adventitious agent detection, and many other applications in the fields of genomics research, drug R&D, and biologics testings. Leveraged with extensive experience in QPCR assay design, validation, and testing, and equipped with the state-of-art ABI 7900 HT Sequence Detection System, We offer a variety of Real Time PCR services to clients around the world.

Real-Time PCR Services

The following real-time QPCR services are offered based upon applications.

Services Descriptions
Copy Number Analysis Target gene specific and genomic DNA assays are used to determine the copy number of a specific gene in cell banks.
Biodistribution Studies Sensitive and target-specific QPCR assays can be custom designed and validated to assess the presence/absence of gene therapy vectors in tissues collected in a preclinical animal safety study.
Residual DNA Testing Sensitive and target-specific QPCR assays are routinely used to detect residual host cell DNA in biological products derived from cell substrates (E. Coli, CHO, Yeast, Human, etc). Residual DNA Testings is also commonly used for cleaning validation.
Gene Expression Analysis The expression levels of specific genes, Small RNA, Micro RNA, and any RNA species of different samples are analyzed using Relative Quantitation Assays predesigned by Applied Biosystems Inc. or custom designed.
TaqMan SNP Genotyping Single Nucleotide Polymorphisms (SNPs) are assessed using TaqMan Allelic Discrimination assays predesigned by Applied Biosystems Inc. or custom designed using ABI’s SNP Genotyping Assay Design Software.
Adventitious Agent Testing The presence/absence of adventitious agents, such as virus and bacteria, is determined with Plus/Minus assays that include target specific PCR primers and probes.
Custom Assay Design and Validation Target specific assays of any species are routinely designed, qualified or validated.
Validated Assay Transfer and Testing Technology transfer of clients’ validated QPCR assays  for the purpose of testing a large number of samples in a cost-efficient way.
QPCR Assay Types

Both Real-Time PCR or End-Point PCR assays are routinely performed at Sunomix Biosciences.

Type Descriptions
Absolute Quantitation Assay Real-time PCR assay with absolute quantitation determined via a Standard Curve.
Relative Quantitation Assay Real-time PCR assay with relative quantitation determined as fold-chage relative to a calibrator or normalizing sample.
Allelic Discrimination Assay End-point PCR assay used to determine the genotype (homozygotes or heterozygotes) of samples.
Plus/Minus Assay End-point PCR assay used to determine the presence or absence of a specific target sequence in a sample.
QPCR Services By Quality Requirements

Sunomix Biosciences offers QPCR services under GLP and GMP regulations promulgated by US FDA (CFR 21 Part 58, Part 210 & 211 and Part 11), European, Japanese, as well as other regulatory agencies worldwide, as applicable to laboratory services provided. We have extensive experience working with clients around the world.

Type Descriptions
Research Grade QPCR DNA sequencing is performed using the same methods as GLP/GMP DNA sequencing, but with less documentation and quality unit involvement.
GLP Grade QPCR DNA sequencing is performed for the purpose of regulatory submission under Good Laboratory Practices (GLP) promulgated by US FDA (21 CFR Part 58 and Part 11), European, Japanese, and other regulatory agencies worldwide.
GMP Grade QPCR DNA sequencing is performed for the purpose of regulatory submission under current Good Manufacturing Practices (GMP) promulgated by US FDA (21 CFR Part 210 & 211 and Part 11), European, Japanese, and other regulatory agencies worldwide.

To find out more details about this custom service, Please request a quote today.