Harlan Laboratories GenScreen™ has
developed the Harlan Laboratories GenCheck™ Genotyping program to
provide high throughput molecular technologies in a format that
clients can easily integrate into their busy laboratories. Standard
PCR is commonly employed for determining the presence or absence of
a gene of interest in transgenic or knockout colonies. We utilize high quality, high fidelity Taq
polymerase from Applied Biosystems to drive PCR in our genotyping
applications. Every sample is run with an internal PCR control to
ensure that there are no
inhibitors of PCR that could lead to inaccuracies in genotyping. In
addition, to maximize throughput and sensitivity of detection,
Harlan Laboratories GenScreen™ employs a variety of technologies,
including Lab-on-a-Chip technology from Caliper (LabChip90) and
fluorescence based PCR.
Quantitative PCR (QPCR) is typically
employed for transgenic and knockout zygosity testing. This
approach is used to measure a fluorescent signal after every PCR
cycle, allowing for analysis of data during the logarithmic phase.
This application utilizes an endogenous single copy gene for data
normalization required to measure the relative copy number of the
gene of interest. Taqman® chemistry and minor groove binding (MGB)
probes are used to ensure success for both easy and challenging
target sequences. Genotyping determinations are made using
validated analytical tools which include comparative
analysis.
SNP
analysis is provided by Harlan Laboratories GenScreen™ for
genotyping applications when a specific point mutation or single
nucleotide polymorphism (SNP) is identified. Both induced and
spontaneous point mutations are measured using Taqman® chemistries.
They are measured in a high throughput manner using Applied
Biosystems 79000HT real time PCR machines which are capable of
generating upwards of 10,000 data points daily. In addition, we
maintain a library of species and strain-specific assays employed
in commonly used research models, including
Leprfa, Leprdb,
Lepob, Prkdcscid,
Foxn1nu and Pdeb6rd1. Custom
assay development is also available, allowing clients to genotype
any mutation of interest.