Here is the link to the paper:
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4276293/
Friday, October 9, 2015
Next Gen Sequencing
The standard protocol for DNA Barcoding is to perform PCR amplifications followed by Sanger sequencing. Sanger sequencing uses dideoxynucleotides and normal nucleotides from DNA. The writers of the paper I read this week propose a new method claiming that its more effective, faster, and cheaper than the standard methods used today. This "next gen" method is called 454 pyrosequencing. this method uses tags that are attached to the primers. The tags are set of oligonucleotides with a known sequence. To prove their statement they performed side by side experiments using both the standard method and the "next gen" method to generate barcodes for 190 species of Lepidoptera. The results of this experiment were that the 454 pyrosequencing method outperformed the standard method. the "next gen" method produced 189 complete barcodes after only 1/8 of a complete run compared to the standard Sanger sequencing only producing 127 complete barcodes. The authors I feel have proved their point. This new method is more accurate and produces results at a much faster rate. This method, in my opinion, should be considered to be the new standard, because with the amount of time and resources saved it could save the barcoding community some money.
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It is cool to see that we are constantly making improvements to modern methods of DNA Barcoding and sequencing. Maybe in a year or two, this method might be outdated and an even better method might arise that's exponentially more efficient.
ReplyDeleteIs research with this more effective method fairly new or is it starting to be used more often?
ReplyDeleteThough this method does have its advantages such as greater flexibility with primer placement, less time then Sanger sequencing with more sensitivity it does have some disadvantages. Some of these disadvantages include that the read data from the software is read in "flow space" instead of "nucleotide space" which essentially means that it considers light intensity measurements instead of the predicted read sequences. It also has issues with repeat regions because it reads such short regions, 300-500 nucleotides. Though it does have some kinks I think eventually combined with other sequencing platforms this has much potential in many scientific fields.
ReplyDeleteGood points, Sarah Long.
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