The GSF requires that total RNA be submitted for mRNA and small RNA sequence analysis. It is recommended that total RNA be purified using the TRIzol® Reagent with Phase Lock Gel™ Heavy procedure (Trizol/Phase Lock Gel Protocol). If retention of miRNA is not required, the Qiagen RNeasy or other comparable isolation kit is acceptable. DNase treatment of RNA samples is strongly recommended to ensure accurate RNA quantitation. Submit RNA samples in nuclease-free H2O at the following concentrations:
The GSF requires that all RNA samples submitted for sequence analysis be assayed spectrophotometrically. All RNA samples submitted will be subjected to an Agilent 2100 Bioanalyzer quality control run. Submit samples in 1.5 -1.7ml microcentrifuge tubes with the name clearly labeled on the top of the tube with permanent marker as it is written on the order form. Please limit sample names to eight characters.
The GSF recommends that genomic DNA (gDNA) submitted for sequence analysis be purified using the QIAamp DNA kit or comparable gDNA isolation kits. RNase treatment of gDNA is required. Submit gDNA samples in nuclease-free H2O or reduced EDTA TE buffer (10mM Tris, 0.1mM EDTA pH 8.0). Single ChIP enriched DNA or control ChIP DNA should be submitted in nuclease-free H2O. Submit either source DNA as follows:
Submit samples in 1.5 -1.7ml microcentrifuge tubes with the name clearly labeled on the top of the tube with permanent marker as it is written on the order form. Please limit sample names to eight characters.
1. Libraries must be prepared using the TruSeq Library prepartion kits or other Illumina kits compatible with the TruSeq SBS sequencing chemistry. For custom library constructs contact the GSF before submission.
2. Investigator prepared libraries must be submitted at 4nM concentration. Download the following conversion tool for converting ng/ul to nM: nM Conversion Calculator
3. When multiplexing, indiviual library concentrations must be adjusted to 4nM prior to pooling.
4. Accurate quantitation of library concentartion is critical for flow cell clustering. It is strongly recommended that qPCR be used for library quantitation.
5. It is recommended that an Agilent Bioanalyzer 2100 quality control run be performed on the library preparations to ensure proper sizing of library.
6. Please provide 15 ul - 20 ul of 4nM libraries in 1.5 -1.7ml microcentrifuge tubes with the name clearly labeled on the top of the tube with permanent marker as it is written on the order form. Please limit sample names to eight characters.
Hand delivery of samples can be made between 9:00 am - 4:30 pm Monday - Friday at the GSF located in RM# 1015 Hemenway, University of Kansas Medical Center. Dry ice shipments of samples, including completed order form, may be shipped overnight to:Genomics Core1015 Hemenway University of Kansas Medical CenterReceiving Dock2106 Olathe Blvd.Kansas City, Kansas 66160
For further information on the Genome Sequencing Facility browse the rest of our web site. Please feel free to call if you have any questions or require directions to the facility.
Phone: (913) 588-7127
Fax: (913) 588-7131