GENOTYPING BY SANGER SEQUENCING
ACGT offers a comprehensive genotyping service by Sanger DNA sequencing to screen variants based on the nucleotide sequence. SNPs in any genome are detected by the PCR amplification of the target region and DNA sequencing the PCR product. PCR product subcloning and screening followed by DNA sequencing is an added service we provide for an increased reliability in detecting heterozygous bases, or for the analysis of rare (low frequency) SNPs. DNA sequencing by Sanger is one of the most definitive methods available for allele identification and genotyping, and is currently regarded as the gold standard in genotyping.
Genotyping by DNA Sequencing Service Description:
- Complete study design and support capabilities
- Design and synthesis of PCR amplification primers and sequencing primers
- Optimization and verification of PCR amplification conditions for new and pre-existing strategies
- Isolation of genomic DNA or total RNA and cDNA synthesis
- PCR amplification and DNA sequencing
- PCR product subcloning/screening followed by DNA sequencing (optional)
- Sequence data filtering and trimming based on Phred 20 to 40 base call quality
- Manual verification of automated base calls in sequence data
- Sequence data comparison analysis between samples and reference sequence
- Extensive troubleshooting experience and technical support