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==Transposon Complex Assembly== Meant to assemble 20 different transposon-Tn5 complexes; with each containing the Tn5 mosaic end sequence at its 3' end to a 5'-phosphorylated 19-bp mosaic end complementary oligonucleotide. Final stock concentration: '''100uM'''. 8 of the oligonucleotides had ends compatible with the P5 Illumina sequencing end, and the other 12 had the adaptor for the P7 side. #Mix Tn5 mosaic ends and complementary oligonucleotides in annealing buffer ##Annealing buffer: 10mM Tris-HCl, 1mM EDTA, 25 mM NaCl, pH 8.0 #Anneal using a themocycler with following parameters: ##95C for 5 min ##decrease to 25C at -0.1C/s #Transposons were then individually mixed at a 1:1 molar ratio with EZ-Tn5 at final stock concentration of 12.5 uM #Incubate at 37C for 1 hour #Assess quality using an 8% TBE gel #Generate 96 transposome complexes ##aliquot, in a 1:1 molar ratio, 8 i5 and 12 i7 transposomes into columns 1-12 and rows A-H, respectively of a 96 well plate ##Store at -20C #Working stock is 2.5 uM and can be stored at -20C
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