Real-time PCR

Real-Time PCR (RT PCR), or 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, Avance Biosciences offers a variety of real-time PCR services to clients around the world.

Real-Time PCR Services

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

Services Descriptions
Copy Number Analysis Gene specific and genomic DNA assays are used to determine the copy number of a specific gene in a cell bank.
Biodistribution Studies Sensitive and target-specific QPCR assays are custom designed and validated in order to assess the presence or 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 at Avance Biosciences to detect residual host cell DNA in biological products derived from cell substrates (E. Coli, CHO, Yeast, Human, etc). Residual DNA test 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 assays developed by Avance Biosciences.
SNP Genotyping by Real-Time qPCR Avance Biosciences assesses single-nucleotide polymorphisms (SNPs) by conducting allelic discrimination assays with TaqMan® probes. Assays may be predesigned by Applied Biosystems Inc. or custom designed using ABI’s SNP Genotyping Assay Design Software.
Adventitious Agent Testing The presence or absence of adventitious agents, such as viruses 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 at Avance Biosciences.
Validated Assay Transfer and Testing Technology transfer of clients’ validated QPCR assays to Avance Biosciences 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 Avance Biosciences.

Type Descriptions
Absolute Quantitation Assay Real-time PCR assay with absolute quantitation determined via a Standard Curve.
Relative Quantitation Assay This real-time PCR assay determines the fold-changes of the expression level of a gene 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.

Compliance

Avance Biosciences offers QPCR services according to GLP and GMP regulations promulgated by the US FDA (CFR 21 Part 58, Part 210 & 211 and Part 11), European, Japanese, as well as other international regulatory agencies, as applicable to the 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 the 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 the US FDA (21 CFR Part 210 & 211 and Part 11), European, Japanese, and other international regulatory agencies.

TaqMan® is a registered trademark of Roche.