2012 © Institute for Molecular Medicine Finland FIMM
DNA / RNA requirements
DNA
- Sequenom / Roche LightCycler
- Ideal concentration is 10 ng/µl (also 5 ng/µl is possible) and the amount needed for one multiplex reaction is 20 ng. DNA dilutions should be made in water
- Provide the samples in skirted 96-well plates
- Purify DNA using commercial kits, quantitate it using either UV spectrophotometer or Picogreen.
- To achieve optimal resources for quality control procedures, we recommend plating 90 unique samples and two duplicate samples per plate. The rest four wells can be left empty and are filled with two CEPH-control samples and two water controls by us.
- Illumina
- Minimun concentration is 50 ng/µl, all samples within a project should be in same concentration
- Total amount of DNA varies according to application, e.g. 200 ng for Infinium genotyping, 0.5-1 ug for methylation analysis, please ask us for details
- In case / control projects you should consider duplicating one or more samples per 96-well plate for quality control
- Affymetrix
- Optimal concentration for samples is 50 ng/ul. E.g. for Genome-Wide Human SNP Array 6.0 chip you need 500 ng of DNA per sample
- Microsatellite genotyping
- A whole genome scan with Linkage Mapping Set v.2.5 - MD10 uses approximately 5 ug of DNA, optimal concentration 5-50 ng/ ul, use same concentration for all samples
- For fine mapping projects please enquire us
- All plates should have at least 3 empty wells for control samples
RNA
- Illumina
- We accept total RNA extracted by standard methods (from non-infectious sources only). RNA should be stored in RNase free tubes or plates (make sure they are sealed properly). The samples can be diluted in good quality RNase free water or TE-buffer (10 mM Tris-HCl, 1mM EDTA). RNA should be of good and uniform quality, 260/280 value between 1.7-2.1 and RIN value >8. If you send samples by courier use dry ice and make sure that they do not thaw during shipment.
- Minimum amount of total RNA is 50-500 ng per reaction, diluted in maximum volume of 11 ul. You should add at least 5 ul extra volume for QC steps.