The gold standard method to identify induced mutations at the target locus entails amplifying the target region by PCR, cloning the amplicon into a vector, followed by Sanger sequencing of the amplicon directly. Each vector should carry only one gene copy, which generates a clean trace in the chromatogram after sequencing. This approach reveals both the frequency and type of mutation at the target locus. It is commonly used to perform screening on clonal cell lines to confirm and validate the desired mutation.
Our experimental setup of Sanger sequencing for mutation screening will design according the gene accession number or rs number provided by the customers. The rs number is an accession number used by researchers and databases to refer to specific SNPs. If the rs number is not available, or customers are providing the primer design(s) to be used in the PCR service, proviso fees may be applicable depending on project requirements. Under this condition, we will offer a custom service work plan to be reviewed and accepted by the customer before confirming the inquiry.
Our regular amplification screening services before Sanger sequencing is now coupled with sample preparation services, allowing you to get the results in the shortest turnaround time, so that you are able to focus in the data analysis and decide the next move of your research experiments.
To maximise success for each sample preparation service, we advise our customers on how to collect and submit the samples within the acceptable terms and conditions. Such information also available in each of the respective order forms. Minimum setup fee will apply when amplification fails due to bad quality of sample that did not meet the prescribed terms and conditions.
Experimental Setup: Genotyping/ Mutation & Variant Screening using qPCR.
PCR Amplification Screening using existing MBS ID
Note: gBlocks™ is the registered trademark from Integrated DNA Technologies (IDT)