Govt. Exams
Entrance Exams
Multiplex PCR amplifies multiple target sequences in a single reaction tube using multiple primer pairs, enabling detection of several genetic markers, genes, or pathogens simultaneously.
qPCR continuously monitors the accumulation of PCR products in real-time using fluorescent reporters (SYBR Green or TaqMan probes), allowing quantification of the target DNA.
rbcL (ribulose-1,5-bisphosphate carboxylase) and matK are standard DNA barcodes for plant identification due to appropriate evolutionary rates and amplification efficiency.
Adapters are short synthetic DNA sequences ligated to fragmented DNA ends, enabling cluster generation, primer binding, and sequencing on NGS platforms.
The PAM sequence (typically NGG for Streptococcus pyogenes Cas9) is recognized by the Cas9-gRNA complex to identify and cleave target DNA at specific locations.
Long-range PCR uses high-fidelity polymerases (like Pfu or Phusion), extended extension times, and optimized Mg²⁺ concentrations to amplify fragments >10 kb.
ddPCR partitions the sample into thousands of water-in-oil emulsion droplets, allowing independent amplification and enabling absolute quantification without standard curves.
WGA using SISPA allows amplification of entire genomes from minimal starting material by using random primers and is useful for viral and microbial genomics.
Multiplex PCR requires careful primer design to prevent primer-primer interactions, dimer formation, and ensure specific amplification of all target sequences simultaneously.
Taq polymerase lacks 3' to 5' exonuclease (proofreading) activity, resulting in ~1 error per 10,000 bases. Pfu has proofreading activity, reducing errors to ~1 per 10 million bases.
