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==NexteraXT== *Single nuclei cDNA (generated on C1 by Rui 05242014) transferred to new 96-well plate using EpiMotion *cDNA for single nuclei was used undiluted for NexteraXT: ''Nextera Tagmentation'' ::{| {{table}} |- |||||||||||<u>Volume</u> |- |Tagment DNA Buffer||||||||||2.5ul |- |Amplification Tagment Mix||||||||||1.25ul |- |Diluted Sample||||||||||1.25ul |- |Total||||||||||5ul |- |} ::*Vortex and spin ::*Incubate 55C 10 min ::*Hold 10C ::*Once sample reaches 10C add 1.25ul of NT buffer in block ::*Vortex and spin ''Nextera Amplification'' ::{| {{table}} |- |||||||||||<u>Volume</u> |- |Tagmented Sample||||||||||6.25ul |- |Nextera PCR Mix||||||||||3.75ul |- |S501 Primer||||||||||1.25ul |- |N7id(1-24) Primer||||||||||1.25ul |- |Total||||||||||10ul |- |} ::''Mix and centrifuge'' ::{| {{table}} |- |72C 3 min|||||||||| |- |95C 30s|||||||||| |- |95C 10s -> 55C 30s -> 72C 60s||||||||||12x |- |72C, 5min |- |} *Combined samples 1-20 (group 1 with N7 ids 1-20) *Combined samples 21-40 (group 2 with N7 ids 1-20) *Combined tube controls *Beads purified (0.8x) twice *Resuspended Group 1 and Group 2 in 50ul; resuspended tube controls in 10ul *Ran 1ul on gel: [[File:07022014_PCR1_Gel1.jpg]]
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