{"id":646,"date":"2013-05-13T01:27:00","date_gmt":"2013-05-13T01:27:00","guid":{"rendered":"http:\/\/suigenerisbrewing.com\/index.php\/2013\/05\/13\/identifying-wild-yeast-part-deux\/"},"modified":"2017-09-15T18:50:20","modified_gmt":"2017-09-15T18:50:20","slug":"identifying-wild-yeast-part-deux","status":"publish","type":"post","link":"https:\/\/suigenerisbrewing.com\/index.php\/2013\/05\/13\/identifying-wild-yeast-part-deux\/","title":{"rendered":"Identifying Wild Yeast Part Deux"},"content":{"rendered":"\n\n\n\n
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From XKCD.com<\/a><\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n

So my first go at identifying wild yeast using ITS sequencing did not work as well as I had hoped<\/a>. Of course, after setting up the system I learned that the method I was using was best for non-yeast fungi, rather than for yeast. Luckily one of my few readers, Sam – author of eurekabrewing<\/a> – came to the rescue with a few papers showing better ways to do it.<\/p>\n

So I’ve revamped the plan and will use a slight variation of my old method. Previously I was trying to sequence the regions of the genome between pieces of the ribosome (full explanation of the method and what a ribosome is can be found here<\/a>). Instead, I’m going to sequence part of the 26s portion of the ribosome (26S rRNA). The specific part I’m amplifying has been used successfully to identify yeast in the past, including the very species of yeast we’re likely to find in a wild brew1-3<\/sup>.<\/p>\n

As always, the meat is below the fold…<\/p>\n

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The Method<\/h3>\n<\/div>\n

So the method is easy – its just a minor adjustment to the old method<\/a>. The only thing that is different is the primers used to amplify the DNA – instead of using the ITS1 & ITS4 primers, we instead use the NS1 and NS4 primers:<\/p>\n