GENOMIC DNA SAMPLE REQUIREMENTS

GENOMIC DNA SAMPLE REQUIREMENTS
WHAT WE REQUIRE
Samples from each individual to be sequenced must contain at least 3ug of high molecular weight
(>50kb) DNA at a concentration >30ng/uL. DNA must be purified, of high quality, and free of non-target
contaminant nucleic acid (RNA, plastid/bacterial DNA). DNA must also be normalized to have equal
concentrations across each sample. To meet these requirements, please use the following instructions.
EXTRACT DNA
We recommend using Qiagen’s DNeasy
Blood & Tissue and DNeasy Plant kits to isolate
genomic DNA.
Genomic DNAs prepared using these kits routinely
generate high quality RAD libraries and sequence
data. DNA sample inputs are critical to the success
of the RAD procedure; please consider this if you
plan to use a different type of kit or homemade
protocol. Feel free to contact the lab staff
([email protected]) prior to DNA isolation with
questions regarding extraction methods.
QUANTIFY
Samples from each individual to be sequenced
must contain at least 3ug of high molecular
weight (>50kb) DNA at a concentration >30ng/
uL. Because DNA quantification determined
using spectrophotometry can vary between labs,
overestimate DNA concentration, and is sensitive to
DNA purity, we recommend that DNA concentration
be assessed by fluorometry. We recommend using
the Qubit Quant iT Assay system (Invitrogen).
CHECK QUALITY
We ask that you visualize your samples on an
agarose gel to check for degradation. Additionally,
we highly recommend using a semi-quantitative
ladder to compare measured concentrations with
the gel image to verify accuracy of quantification
measurements.
NORMALIZE
RAD sequencing on two or more samples requires
DNA concentration to be normalized (equivalent
concentrations) across each sample to be analyzed.
Samples must be normalized to a concentration
of 30ng/uL or greater. It is highly recommended
that you run a gel with normalized material to
visually check that samples appear to be of equal
concentrations, as samples that do not adhere to
this standard will not be processed.
SEND SAMPLE INPUT
AND QUALITY DOCUMENTATION
Send an electronic copy of your gel image(s)
and sample manifests to [email protected]
for quality approval before shipping your samples.
Wait for approval from lab staff before continuing
with shipment.
PREPARE SAMPLES FOR SHIPMENT
• 1-7 samples: Samples should be shipped in
1.5-2mL tubes, each clearly labeled with a sample
name and shipment date. Wrap the tops of your tubes
in parafilm and place them in a plastic conical or other
hard container stuffed with tissue to prevent damage
during transport.
• >7 samples: Samples should be shipped in 96-well
plates, each clearly labeled with a plate name and
shipment date. Please seal the plates in plastic bags
and pad them with bubble wrap or paper towels to
prevent damage during transport. Samples shipped in
individual tubes will be returned.
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