How many nucleotides does a DNA probe need to only mismatch 119 times in a 2 bil
ID: 65317 • Letter: H
Question
How many nucleotides does a DNA probe need to only mismatch 119 times in a 2 billion base par air genome?
Explanation / Answer
Method Read length Accuracy (single read not consensus) Reads per run Time per run Cost per 1 million bases (in US$) Advantages Disadvantages Single-molecule real-time sequencing (Pacific Biosciences) 10,000 bp to 15,000 bp avg (14,000 bp N50); maximum read length >40,000 bases[61][62][63] 87% single-read accuracy[64] 50,000 per SMRT cell, or 500–1000 megabases[65][66] 30 minutes to 4 hours[67] $0.13–$0.60 Longest read length. Fast. Detects 4mC, 5mC, 6mA.[68] Moderate throughput. Equipment can be very expensive. Ion semiconductor (Ion Torrent sequencing) up to 400 bp 98% up to 80 million 2 hours $1 Less expensive equipment. Fast. Homopolymer errors. Pyrosequencing (454) 700 bp 99.9% 2 million 24 hours $10 Long read size. Fast. Runs are expensive. Homopolymer errors. Sequencing by synthesis (Illumina) 50 to 300 bp 99.9% (Phred30) up to 6 billion (TruSeq paired-end) 1 to 11 days, depending upon sequencer and specified read length[69] $0.05 to $0.15 Potential for high sequence yield, depending upon sequencer model and desired application. Equipment can be very expensive. Requires high concentrations of DNA. Sequencing by ligation (SOLiD sequencing) 50+35 or 50+50 bp 99.9% 1.2 to 1.4 billion 1 to 2 weeks $0.13 Low cost per base. Slower than other methods. Has issues sequencing palindromic sequences.[70] Chain termination (Sanger sequencing) 400 to 900 bp 99.9% N/A 20 minutes to 3 hours $2400 Long individual reads. Useful for many applications. More expensive and impractical for larger sequencing projects. This method also requires the time consuming step of plasmid cloning or PCR.