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Sequencing TechniquesWhat do the terms ‘assembly’ and ‘mapping’ mean in the context of sequencing experiments? How...

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Biology

Sequencing TechniquesWhat do the terms ‘assembly’ and ‘mapping’mean in the context of sequencing experiments? How do Sanger andIllumina sequencing work (just in general terms)What samplepreparation steps are generally shared among Next GenerationSequencing technologies?

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ANSWER Assembly means construction of longer sequence from smaller sequence readsPowerful computer algorithms are utilized to piece the resulting sequence reads back together into longer continuous stretches of sequence contigs a process known as de novo assembly For correct assembly it is important that there is sufficient overlap between the sequence reads at each position in the genome which requires high sequencing coverage or read depth Naturally for longer sequence reads more overlap can be expected reducing the required raw read depth Usually longer fragments several hundred base pairs are sequenced from both ends pairedend sequencing to provide additional information on correct read placement in the assembly The mapping is the process of comparing each one of the reads with the reference genome We will obtain one alignment or more between each read and the genomeWhen studying an organism sequence with a reference sequence it is possible to infer which transcripts are expressed by mapping the reads to the reference genome genome mapping or transcriptome transcriptome mapping Mapping reads to the genome requires no knowledge of the set of transcribed regions or the way in which exons are spliced together This approach allows the discovery of new unannotated transcripts ANSWER Sanger sequencing also known as the chain termination method is a method for determining the nucleotide    See Answer
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