Matheo Analyzer 3.2 Keygen
Saija Grzegorek
In the next PCR, Illumina index adapters were added to all amplified tiles. Each PCR consisted of: 20 µL Phusion High-Fidelity PCR Master Mix with HF Buffer (NEB), 2 µL 10 µM i5 indexing adapter, 2 µL 10 µM i7 indexing adapter, 1 µL 1:10 diluted PCR product, 15 µL nuclease-free water. Tiles were amplified using the following program: 98 C 30 s, 7 cycles of 98 C 15 s, 65 C 30 s, 72 C 120 s, followed by 72 C 7 min and hold at 4 C. An equal volume of each PCR was then pooled and 100 µL were used for gel extraction from a 4% E-Gel EX Agarose Gel (Invitrogen). The fragment size and quality of the extracted DNA were tested using a 2100 Bioanalyzer system (Agilent), and DNA concentration was determined using Qubit (ThermoFisher). Finally, libraries were paired-end sequenced using an Illumina NextSeq 550.
The authors thank Sofie V. Nielsen for helpful conversation on experimental procedures, and Amal Al-Chaer for assistance with the Bioanalyzer system and Illumina sequencing. We acknowledge the use of computing resources at the core facility for biocomputing at the Department of Biology, University of Copenhagen.
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