Illumina HiSeq 2500 paired-end Sequencing of KAS20000465 (KAE20000270)
missing
We lysed 50,000 cells with 50 microliter of Lysis buffer 1-2mM Tris-Cl pH 7.4, 1-2mM NaCl, 3 mM MgCl2, 0.1-2NP-40, 0.1-2Tween-20, and 0.01-2Triton X-1-2 and incubated them for 5 min on ice . Following centrifuging at 1-200 g for 1-2min at 4 degree Celsius, we resuspended the nuclei pellets with 50 microliter of transposition mix 25 microliter 2X TD buffer 20 mM Tris-HCl pH 7.6, 1-2mM MgCl2, and 20% Dimethyl Formamide in sterile water and 2.5 microliter transposase 1-2 nM final, 1-25 microliter PBS, 0.01-2Triton X-1-2, 0.1-2Tween-20, and 5 microliter H2O and incubated the mixture with shaking for 1-2 at 55 degree Celsius in a thermomixer. We purified transposed DNA using DNA Clean and the Concentrator-5 Kit Zymo Research. We amplified the transposed DNA using the NEBNext High-Fidelity 2X PCR Master Mix for 1-2cycles.
KOBIC
KAE20000270
Pohang University of Science and Technology
DRX601991