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=Low-Input Transposase-Based Library Preparation= ==Klenow Extension Test== * Ran gel from yesterday's experiment * Loaded as: Low-Mass Ladder, 100 pg, 10 ng, NTC [[File:02182011_Low_Input_Klenow.jpg|600px]] * Both libraries seemed to work well * '''Klenow appears able to replace the Nextera extension step''' * Maybe wait a bit longer in next protocol? Waited 20 minutes, but have the appearance of lower efficiency. ** We could probably increase this by waiting longer ==Bead Capture Experimental Plan== * Repeat Klenow protocol from yesterday * Instead of going to PCR step, add TdT and TdT buffer with Biotin-labeled dATPs * Capture DNA with beads, wash, amplify * Check supernatant for DNA after Nextera PCR; run gel on supernatant ==TdT Extension Test== * TdT is not very efficient with double-stranded blunt-end DNA * Options: ** Denature before labeling; KOH - HCl denaturing? ** Increase incubation time suggested by protocols ** Found one dsDNA labeling protocol, but states that it is inefficient
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