Biotechnology

NGS and allele-specific digital PCR can detect rare mutations (ctDNA) with high sensitivity; RFLP/DGGE are less sensitive for point mutation detection.

ddPCR partitions samples into independent reactions, allowing Poisson distribution analysis for absolute quantification without reference standards.

Inverse PCR amplifies unknown DNA sequences flanking a known region by using outward-facing primers on a circularized DNA template, useful for genome walking and transposon mapping.

MGB sequences bind in the minor groove of DNA, stabilizing the probe-DNA duplex and increasing its melting temperature, allowing for shorter, more specific probes.

For plant DNA barcoding, rbcL (ribulose-1,5-bisphosphate carboxylase/oxygenase large subunit) and matK (maturase K) are the standard barcode regions recognized by CBOL.

ARMS-PCR uses allele-specific primers that only amplify when the primer sequence perfectly matches the target DNA, making it ideal for detecting specific point mutations.

Competitive PCR uses an internal control (competitor) DNA that is amplified simultaneously with the target using the same primers, allowing relative quantification by comparing product amounts.

HRM analysis detects genetic variations by monitoring the melting temperature (Tm) of PCR products; sequence variations cause different Tm values, visible as distinct melting curves.

GWAS uses Chi-square tests with Bonferroni correction (or FDR correction) to account for multiple hypothesis testing across millions of SNPs and identify genome-wide significant associations.

Inverse PCR uses outward-facing primers on a circularized DNA template to amplify and sequence flanking regions of known sequences, useful for finding regulatory elements.