BioMicroCenter:Sequencing Quality Control: Difference between revisions
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== | == Why is QC Important? == | ||
It is very important to supply us with a reliable measurement of the concentration and quantity of starting material so that we can prepare the sample for sequencing. If a sample is too concentrated the cluster density will be too high and the GA will not be able to detect them properly. If a sample is too dilute the cluster density will be too low and there will not be enough reads for the pipeline to make base calls. Having reliable QC measurements of samples allows us to optimize the cluster density and the number of reads per lane and maximize the quality of data produced. QC can either be run before sample submission and specified upon testing request or can be run by the BMC. We strongly recommend that you allow us to run QC of samples. | |||
Revision as of 12:24, 12 January 2009
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Why is QC Important?
It is very important to supply us with a reliable measurement of the concentration and quantity of starting material so that we can prepare the sample for sequencing. If a sample is too concentrated the cluster density will be too high and the GA will not be able to detect them properly. If a sample is too dilute the cluster density will be too low and there will not be enough reads for the pipeline to make base calls. Having reliable QC measurements of samples allows us to optimize the cluster density and the number of reads per lane and maximize the quality of data produced. QC can either be run before sample submission and specified upon testing request or can be run by the BMC. We strongly recommend that you allow us to run QC of samples.
Below are some of the most common techniques used for QCing samples for sequencing:
- Bioanalyzer - suggested for samples exceeding 10nM although has shown to be less reliable at lower concentrations. Available at the BMC for $10 per sample
- RT-PCR, SYBERgreen assay - coming soon to the BMC!!
- Nanodrop - not recommended for sequencing samples because this method has a detection limit of about 12nM to 22uM and has shown to be less reliable on the lower end of this range.
- Qubit - not tested at the BMC but information about this application can be found at http://openwetware.org/index.php?title=BioMicroCenter:Sequencing_Quality_Control&action=edit&redlink=1
