License: This is an open access protocol distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited
Protocol status: Working
We use this protocol and it's working
Created: November 06, 2023
Last Modified: November 20, 2023
Protocol Integer ID: 90473
Keywords: DNA transport, DNA shipment, packaging DNA, DNA import, DNA export, sample transport
Funders Acknowledgement:
National Institute for Health and care Research
Grant ID: 16/136/111
Abstract
DNA eluted in nuclease-free water or low-TE solution is stable enough to be shipped at ambient temperatures overseas. This characteristic of DNA becomes vital when dealing with logistics and finances of research in low- and middle-income regions with limited access to dedicated shipment carriers and bespoke courier services, not to mention door-to-door delivery delays. Here we describe an efficient and inexpensive method of transporting DNA isolates in a high-throughput format on 96-well PCR microplates.
10. Consignment containers (provided by local courier provider, in this case FedEx)
Before Starting
Before Starting
Ensure that all of the materials listed are available before you begin aliquoting and packaging DNA samples for transport. Contact your local courier provider to obtain consignment containers.
Aliquoting DNA into a 96-well plate
Aliquoting DNA into a 96-well plate
Using filter tips, transfer the required amount of each DNA sample into the 96-well plate (one well per sample).
Record the aliquoted positions on a sheet alongside isolate/sample ID and estimated DNA concentration (in ng/ul) and yield (ng or ug).
Sealing the 96-well plate
Sealing the 96-well plate
Gather the number of 8-cap strips as fully and partly-filled plate columns (Fig. 3a) and place them on a clean, sterile surface, domes facing downwards (Fig. 3b).
Gently line the cap strip along the wells of a single column (domes now facing upwards). Using your thumb, press down on each of the circular domes, one at a time, until they click and lock into the wells (Fig. 4).
Gripping the strip cap applicator as shown, gently roll it back and forth about 10 times along the dome strip in a vertical line (Fig. 5). This will help to lock the caps in with an even amount of pressure across all of them.
Hold the plate up at your eye level and carefully check that all of the strip caps are locked flat into each well (Fig. 6). Loose caps could cause sample leakage and cross-contamination during transport.
Repeat steps 3-7 for each of the other columns filled with samples (Fig. 7).
Securing the 96-well plate for transport
Securing the 96-well plate for transport
Place the sealed plate on a piece of flat sheet cardboard. Using a pencil, trace the outline of the 96-plate (Fig. 8a).
Cut out two such outlines. You will now have two rectangular pieces of cardboard roughly 128 x 86 mm each (Fig. 8b)
Place one piece of cardboard below and the other above such that the plate is sandwiched between them (Fig. 9).
Bind the sandwich together using packaging tapeby winding it tightly around the bundle several times (Fig. 10).
Wrap the taped bundle in a double layer of absorbent tissue (Fig. 11).
Using a large sheet of bubble paper, wrap the bundle within several layers. Next, secure the bubble-wrapped bundle with more packaging tape going all the way around as shown (Fig. 12).
Label the plate bundle as shown below.
Repeat steps 2-13 for each new 96-well plate of samples you intend to ship.
Boxing the secured 96-well plate
Boxing the secured 96-well plate
Carefully place each wrapped plate bundle into the courier container (Fig. 14). If you choose to use a FedEx Small Box, then you can fit a maximum of SIX wrapped 96-plate bundles.
Include a printed paper copy of the sample manifest indicating the positional layout of DNA samples on each plate.
Seal the FedEx box by peeling off the adhesive strip, folding the flaps and firmly pressing down on the upper lip of the box (Fig. 15).
Place the box into a UN3373 Pak (Fig. 16), also provided by your local courier. Upon sealing the bag, record the weight of the entire package.
Contact your chosen courier to arrange payment and a collection date/time.
Place the package in the freezer -20 °C until it is ready to be collected for transport.