Biotechnology

PCR follows exponential amplification: Final copies = Initial copies × 2ⁿ, where n = number of cycles. 2 × 2³⁰ = 2 × 1.07 × 10⁹ ≈ 2 × 10⁹ copies.

RAPD-PCR uses random primers to generate DNA fingerprints for assessing genetic diversity, distinguishing crop varieties, and parentage analysis in breeding programs.

Stem-loop RT-PCR uses a stem-loop primer structure that specifically binds to mature miRNAs, making it ideal for miRNA detection and quantification in diagnostic applications.

Site-directed mutagenesis uses primers with mismatches at the desired mutation site. During PCR extension, the mutated sequence is copied, incorporating the specific change.

AAVs are preferred due to minimal immunogenic response, ability to target specific tissues, and safe integration properties, despite a packaging limit of ~4.7 kb.

Digital PCR partitions the sample into thousands of individual reactions (droplets), allowing absolute quantification by counting positive and negative partitions without requiring standard curves.

COI (cytochrome c oxidase I) is the standard DNA barcode for animal identification due to its high interspecific variation and low intraspecific variation.

Hot start Taq polymerase is inactive at room temperature and becomes activated only at high temperatures (95°C), preventing unwanted non-specific amplification during setup.

NGS provides massively parallel sequencing, generating millions of reads simultaneously, offering significantly lower cost per base and much higher throughput compared to Sanger sequencing.

Annealing temperature is typically 3-5°C below the Tm (melting temperature). For 50% GC content, Tm ≈ 55-60°C, making this the appropriate annealing range.