TITLE: Preparing Microbiome Samples for Cryo Shipment
AUTHORS: Alicia Rich

[DESCRIPTION]
<div class = "text-blocks"><div class = "text-block">This is a generic protocol written for any collaborators or contributing institutions sending samples to the Rich Lab at Otterbein University for microbiome or other genetic analyses. The protocol is written for samples that have already been frozen. They may be stored in any stabilizing solution or dried.</div></div>

[STEPS]
?. Keep all samples on cooling blocks or ice during these procedures.
on ice
?. Label sterile, RNase/DNase-free tubes for each sample that you will be transferring. You should label them using a thin-tip permanent marker, and copy the information on the previous tube. If that information is not visible, add your initials and the date. Try to make out the sample ID # and add a ? if necessary.
Make sure there are labels on the side and the top of each tube. This ensures that the sample will still be labeled if we lose the cap and that you will be able to easily see the sample number on the tops of tubes looking down into the storage box.
Be careful not to touch the inside of the lids or the tubes, even with your gloved hands. Also try not to touch your face or hair while wearing the gloves, and don't cough or lean over the open tubes. If you need to touch something that might contaminate your gloves, then change out a new pair.
?. Use the sterile tweezers to carefully transfer all of the material from an old tube into the new, fully labeled tube. Again, take care not to touch your finger to the sample or the inside of the lid. Do not use the same tweezers for more than one sample, or if you must, then clean the tweezers between samples with a 10% bleach solution.
?. As soon as you complete one sample, cap both tubes. Put the old tube into a biohazard disposal bag. Put the new tube back into the freezer immediately or keep it on ice.
The most important thing is that you do not transfer genetic material from one sample to the next when you do this. Consider rubbing a few drops of the 10% bleach solution on your gloves between samples or change gloves if you think you inadvertently touched any sample material before you move to the next one.
?. After you finish all of the tubes, carefully arrange them in the sample box that I provided and double check that all of the caps are labeled with the sample #. Put this box into the freezer until shipment.
?. Properly dispose of the biohazard bag with the old tubes. This will depend on your lab's protocols, but you may need to put autoclave tape on the bag, autoclave it for sterilization, and then put it in the trash for disposal. Only materials that may have remaining biological materials on them are considered biohazard waste requiring autoclaving before disposal.
?. Proceed to the steps for sample shipment.
?. Make sure all tubes are securely capped and labeled. Tubes must be placed in a secondary container. For small tubes (2 mL and less), this should be freezer boxes. For large tubes, this can be sealed ziploc bags.
?. Line the inside of a styrofoam cooler with a large, plastic bag. Put this inside a freezer (  if available) overnight to pre-chill it.
-80 °C
Do not add the dry ice yet. Just add the plastic liner as shown here.
?. Put these boxes or bags inside the plastic-lined styrofoam cooler. Make sure you have left enough room for dry ice around the edges and on top. Keep this inside the freezer until immediately before shipment.
?. Make sure all labels are correctly placed on the outer cardboard box.Proper shipping name: "DRY ICE"UN NUMBER: "UN 1845"Class 9 Hazard Label (attached)   Net quantity of dry ice "2 kg"FedEx Express electronic shipping label with shipper and recipient markings on it:        Your name and address as shipper        Recipient name and address:Class 9 Hazard Label (attached) Net quantity of dry ice "2 kg"FedEx Express electronic shipping label with shipper and recipient markings on it:        Your name and address as shipper        Recipient name and address:                       Rich Lab c/o Erin Ulrich                       Department of Biology and Earth Science                       Otterbein University                       1 South Grove Street                       Westerville, OH 43081
?. Assemble the documents that will need to go inside the cardboard box (but outside the styrofoam cooler). That includes the shipment letter from Dr. Rich and the sample manifest (both will be provided).
?. I will schedule a pickup for the package. Soon before, you should place the styrofoam box inside the cardboard box. Close the plastic liner as shown below. Surround it with dry ice and place the styrofoam lid on top.Tape the box as shown in step 5.
Do not seal the lid on. Air must be able to escape the box as the dry ice evaporates.
?. Hand off the package for pickup! Thank you!