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==Background== *I have already made Illumina libraries from the four highest concentrated samples on <http://genome-tech.ucsd.edu/LabNotes/index.php/Chris:LabNotes/SeaMicrobiome/Calendar/2015/2015-2-18>. These four samples will also be used in a comparison test with two Nextera enzymes (original Nextera and supermutant): Concentration CH-S-A 63.2ng/ul CH-S-B 69.2 CH-S-C 83.9 CH-S-D 35.2 *We want to dilute the samples to ~1ng/ul using the following dilutions: Concentration ul Sample ul Water CH-S-A 63.2ng/ul 1 62 <- We will be using only this sample for today's test CH-S-B 69.2 1 68 CH-S-C 83.9 1 82 CH-S-D 35.2 1 34 *These libraries will be tested using the protocol provided by Andrew on <http://genome-tech.ucsd.edu/LabNotes/index.php/Arichard:Protocols/tn5059>. The necessary reagents can be found in one of three places: **Andrew's 4C box **-20C 2nd shelf in Midas -20C box **-20C 2nd shelf in Nextera primer box *The Nextera supermutants are provided in the 50ml tubes labeled Control (the thick handwriting is 25x diluted enzyme) *For this test, we'll need to run several reactions: different dilutions of the enzyme (up to 1x dilution and below) and also include a control tube with the original Nextera enzyme (already diluted 1/50) *Below are the reactions and conditions we'll be testing today: Rxn # Template Enzyme Enzyme Dil 1 CH-S-A (1ng) Nxt Old 1/50 2 "" Nxt SM 1x 3 "" "" 0.75x 4 "" "" 0.50x 5 "" "" 0.25x 6 "" "" 0.1x 7 Water (NTC) Nxt Old 1/50 8 "" Nxt SM 1x
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