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===24th=== * We have 4 libraries that we are trying to catalog: 10/1, 10/3, 10/10, and 10/12. * We are implementing a new system of data storage. Every sample will have an entry on a spreadsheet with links to each experiment page, which will contain all data including gel images, PCR data and any notes or changes to the protocol. This information will be updated in real-time for all experiments going forward. * The clonal rates are high for the latest libraries (70-90%). We will add BST polymerase and dNTPs to the protease reaction in order to fill in gaps left by tagmentation. This should help to prevent the loss of the priming regions added by transposase. We will continue to do Exo- treatment as well, to be safe. * BST has optimal activity at 65 degC, but can be used up to 70 degC. Inactivation is at 80 degC for 20 minutes. * We can add BST after the 15 minute protease treatment, just before the 20 minute protease inactivation at 70 degC. This will prevent the protease from destroying BST before it has had a chance to act.
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