Aquaculture & Marine Biotechnology

• TaqMan Probes in Quantitative Real-Time PCR:
• TaqMan Probe: An oligonucleotide probe dual-labeled with a $5'$ fluorescent reporter dye (e.g., FAM) and a $3'$ quencher dye (e.g., TAMRA).
• During real-time PCR, Taq DNA polymerase's $5' \to 3'$ exonuclease activity cleaves the hybridized probe, releasing the reporter from quenching and generating fluorescence proportional to accumulated amplicon. It is the gold standard for genotyping and SNP detection in real-time PCR.
• Correct Option: Option 1 (genotyping by real-time PCR).

• Invention of Polymerase Chain Reaction (PCR):
• The Polymerase Chain Reaction (PCR) technique was conceived and developed in 1983 by American biochemist Kary B. Mullis at Cetus Corporation.
• PCR allows exponential enzymatic amplification of specific target DNA sequences in vitro using thermostable DNA polymerase, earning Mullis the 1993 Nobel Prize in Chemistry.
• Correct Option: Option 1 (Kary Mullis).

• Determination of Transgene Insertion Sites:
• When a transgene integrates randomly into host genomic DNA, only the internal sequence of the transgene vector is known; the surrounding flanking genomic DNA is unknown.
• Inverse PCR (IPCR):
• Designed specifically to amplify unknown sequences flanking a known core region.
• Genomic DNA is digested with a restriction enzyme cutting outside the known core, diluted to favor self-circularization under low concentration, and ligated into circles.
• Primers oriented in opposite directions (inverted outward from the known sequence) amplify the circularized unknown flanking genomic region.
• nalysis:
• (Inverse PCR): Correct.

Correct option: (1) Inverse PCR