Aquaculture & Marine Biotechnology

• 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

• BAC and YAC Cloning Vectors:
• Bacterial Artificial Chromosomes (BACs): Based on the E. coli fertility ($F$) plasmid, carrying large inserts ($100 - 300\text{ kb}$) at a strictly controlled single copy per host cell.
• Yeast Artificial Chromosomes (YACs): Linear vectors containing yeast centromere (CEN), telomeres (TEL), and autonomous replication sequences (ARS), accommodating inserts up to $1000 - 2000\text{ kb}$ as single-copy linear chromosomes.
• Correct Option: Option 4 (High capacity, single copy vectors).