Experimental Principles (Sequencing/HiSeq2000)

We carry out sequencing with the HiSeq2000. Clusters are synthesized from randomly fragmented DNA or cDNA, and the reversible terminator technique is used to read signals with a high degree of accuracy to determine base sequences.
Sample quality check

For DNA, PicoGreen is used to measure the concentration, and electrophoresis to confirm the size.

For RNA, a bioanalyzer is used for quality evaluation.

Library quality check

The DNA or RNA is fragmented and adaptor ligation is performed.

Cluster formation

DNA is bound to flow cells to form clusters.

Sequencing

Reversible terminators are used to perform single-base extension reactions while images are acquired, following the sequencing-by-synthesis methods. Data are output in FASTQ file format.

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