Download Genome Wide Sequencing

Genome Wide Sequencing
How much DNA is needed and what are the requirements?
gDNA isolated by most standard techniques is acceptable but should be RNase treated and purified and
have a concentration of 100ng/ul. Standard column techniques work well. A 260/280 of >1.7 is
recommended and quantitation should be done on a NanoDrop and Qubit spectrofluorometer.
Can I submit less than 100 ng/ul?
Yes by special arrangements only. Be aware that the core may need to increase the recommended
coverage in that case.
Do I need to know the GC content prior to submission of my bacteria?
Yes, this is very helpful when determining coverage requirements
Do I need to run my DNA out on a gel or use the Bioanalyzer before submission to the Core?
Yes, the core will need to see the size distribution prior to starting the library synthesis. This is because
our library input size needs to be 300-400bp AFTER a fragmentation step. The fragmentation can only be
done if we know the input size distribution.
What is the coverage and read length required?
200 base pair paired end is needed for GWS. Coverage depends on the PI needs and GC content.
Generally 100x coverage is acceptable but be aware the sequencing of repeat regions is not standard for
the Illumina HiSeq due to the sequencing length limit.