SAMPLE PREPARATION

Sample quality directly impacts sequencing quality and the downstream analysis. When you are submitting samples to genomine, it is crucial to follow the below given instructions. This will help us to successfully complete your project.

Input quantity should be determined by Qubit instead of by NanoDrop . If a spectrophotometer (e.g. Nanodrop) is used, we suggest submitting twice the requested amount of sample since this type of measurement is often unreliable.

DNA

DNA samples need to be RNAfree and dissolved in EB buffer or TLE buffer.

Shipping DNA that has been dissolved or re-constituted in TE (10 mM Tris, pH 8, 0.1 mM EDTA) or 10 Mm Tris pH 8.0 with 4°C wet ice packs (blue ice) is the recommended way to ship DNA samples.
There are different methods by which you can ship your DNA. You probably can ship extracted DNA on dry ice. You may also ship pure DNA sample in water or DNA in lyophilized form at ambient temperature  , Saliva samples can be shipped at room temperature.

DNA samples are required to have an OD260/280 ratio between 1.8 and 2.0 and 260/230 = 2.0
For DNA sequencing the ideal concentration of RNA should be around 5-10 µg and no less than 1µg.

Samples not meeting these specifications should be designated as “poor quality” by the customer. These samples will be sequenced at your own risk and billed regardless of the data quality.

RNA

RNA samples need to be DNAfree and should be dissolved in molecular biology grade water (RNAsefree; not DEPC treated).

For most RNA applications, you’ll need to ship total RNA. Clean the surface of all tubes and use nuclease-free tips. Use nuclease-free water for buffer preparation and reconstitution of your total RNA sample.

RNA samples are required to have an OD260/280 ratio between 1.8 and 2.0 and 260/230 = 1.7
RNA samples should be kept in dry ice during shipment.

For RNA sequencing the ideal concentration of RNA should be around 5-10 µg and no less than 200 ng.

Fresh Tissue

If you would like us to extract RNA or DNA from fresh tissue it should be quickly frozen and shipped on dry ice.

Constructed Library

A constructed library is a very stable, double stranded PCR product. It is ideal to have between 2 –10 nM of that library for sequencing. If you’re shipping a library you’ve constructed yourself, reconstitute your library in ~10-50 µl of 10 mM Tris pH 8.0 buffer and use microcentrfuge tubes that are 'low-bind'. At very low concentrations, libraries tend to stick to tube walls.

Packing and Shipping.

Avoid sample labels from being dissolved by solvents and from falling off the tubes. Use a waterproof marker pen to write directly on the tube wall or lid. We recommend you print and stick the label onto the tube wall with clear, adhesive tape completely around the tube.

Please enclose a hard copy of Sample Information form in the shipment. We also recommend you E-mail the Sample Information form directly to us before shipping the samples.

Before you ship please make sure that the sample information in the Sample Information form matches the labels on the tubes.

Place your DNA or RNA sample in a 1.5 or 2 mL screw cap microcentrifuge tube and seal with Parafilm. Pack the tubes in a thermo-stable shipping box. The tubes may get crushed during shipping; we highly recommend placing them in secondary container such as a box with interior racks/holders. We do not recommend using 96-well plates and PCR stripe tubes for shipping your samples.

We recommend the Styrofoam be at least 1.5 inches thick. Fill the box with dry ice or wet ice. 3-4 kg of dry ice should last for at least 48 hours. Ask the courier service how long does it take for your shipment to reach the destination. Find the right size box and fill it with dry ice or wet ice accordingly this also prevents tubes from moving around inside the container. When dry ice or wet ice is not used shredded paper, foam or cotton can be used as stuffing to prevent movement inside the container.

Use the fastest available courier, e.g. FedEx, UPS, DHL or USPS. We recommend selecting Priority Overnight service. The carrier may ask you to fill out a commercial invoice. Label your contents as ‘non-hazardous research sample’. Label the box as ‘temperature sensitive, keep frozen’. Send us the tracking number so we know when to expect your package.

Try to ship on Monday or Tuesday and not close to weekend or public holidays to avoid delay associated with delivering on a weekend and holidays. Shipping during holidays can unnecessarily delay the time it takes for us to unpack and properly store your samples.

Depending on your location, you will ship your samples either to our office in US or India.