DNA

Q: How do I collect and submit blood samples for DNA extraction?

Blood should be collected in purple top vacutainers (EDTA as additive). Store the tube in a 4°C refrigerator and submit to CMG on wet ice within 48hr of blood draw.

Q: How much DNA will I get from my blood samples?

Many factors may affect DNA yield, such blood handling and storage condition, white blood cell count, and etc. Typically, about 300µg of genomic DNA can be extracted from 10ml of whole blood.

Q: How much DNA do you ask for genotyping?

The amount of DNA needed depends on the specific application, project size and platform used. As a rule of thumb, SNP analysis (e.g. TaqMan, SNPstream) usually only requires nanograms of DNA per assay, whereas whole genome chip or methylation analysis (e.g. Illumina Infinium) typically uses micrograms. For certain applications, we also perform additional QC steps before the analysis therefore extra DNA is needed.

Q: Who should I contact if I need further information for my study?

Co-Directors Drs. Mark Bouzyk (DNA) & Carlos Moreno (RNA) or Senior Research Associate Dr. Weining Tang. Please see contact us page

RNA

Q: How much RNA do I need to submit?

Illumina applications require 2 µg of total RNA per sample is best with a minimum concentration of 40 ng/µl.
Affymetrix applications require 11 µg of total RNA per sample with a minimum concentration of 625ng/µl.

Q: What if I am doing laser-capture microdissection?

If you are doing LCM or have other experimental reasons for very small RNA yield, we can perform RNA amplification starting from as little as 50 ng of RNA.

Q: How should I prepare my RNA?

If you are interested in examining microRNAs at a later time, we recommend the miRVana kit from Ambion, but Trizol also recovers large and small RNA species. If you have no interest in microRNAs, other commercial kits such as RNeasy from Qiagen is fine.

Q: How many replicates do I need?

For in vitro cell culture type studies, a minimum of three biologically independent replicates is needed, but four replicates is better statistically. For in vivo animal studies, it is preferable to have six or more animal RNA samples for each group, since there is likely to be greater biological variability. For clinical studies on human samples, a minimum of twelve cases in each group is needed, but more is always better.

Q: Do we accept every project?

No. To ensure best use of our time, investigators money, and an appropriate turn around time for other investigators we reserve the right to decline projects for which we believe our services will not be able to answer the scientific question posed by the experimental design.

Sample Source	DNA	RNA
Whole blood - 10mL	100 - 300µg	30µg
Buffy coat	100 - 300µg	30µg
Serum 1mL	nanograms	nanograms
Tissue 1mg	0.2 - 3µg	1 - 6µg
Urine (1mL)	nano - µg	nanograms
Saliva / oragene - 2mL	100µg	20µg
Cell line - 5 x 106	15 - 20µg	50 - 150µg
FFPE - 5 x 5µm sections	1 - 3µg	1 - 3µg

Instrument	gDNA	FFPE DNA	RNA	FFPE RNA
AB3730 Sanger Sequencing	20 - 50ng	20 - 50ng	N/A	N/A
AB TaqMan Genotyping / Expression	5 - 20ng	5 - 20ng	50ng / assay	50ng / assay
Beckman SNPStream genotyping	2 - 10ng	2 - 10ng	N/A	N/A
Illumina GoldenGate / DASL - genotyping, methylation, & expression	250ng	250ng	200 - 500ng	200 - 500ng
Illumina DirectHyb - expression	N/A	N/A	> 1µg	N/A
Illumina Infinium - genotyping & methylation	400 - 500ng	N/A	N/A	N/A
Affymetrix GeneChips - genotyping, re-sequencing, expression	500ng	N/A	5µg	50 - 100ng

• CRIG - Clinical Research In Genetics
• HIPAA
• HIPAA Revocation
• Informed Consent