Oct 02, 2025
REDI-NET FV-3 FLYING VECTOR STORAGE
- REDI-NET Consortium1
- 1REDI-NET Consortium
- Remote Emerging Disease Intelligence - NETwork (REDI-NET)
Protocol Citation: REDI-NET Consortium 2025. REDI-NET FV-3 FLYING VECTOR STORAGE. protocols.io https://dx.doi.org/10.17504/protocols.io.81wgbkm11gpk/v1
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: June 20, 2025
Last Modified: October 02, 2025
Protocol Integer ID: 220633
Keywords: STORAGE PROCEDURE FOR UNTREATED SAMPLE, STORAGE PROCEDURE FOR TOTAL NUCLEIC ACID, flying vector, storage, flying vector storage, vector storage, vector, redi
Funders Acknowledgements:
USAMRAA
Grant ID: W81XWH-21-C-0001
USAMRAA
Grant ID: W81XWH-22-C-0093
USAMRAA
Grant ID: HT9425-23-C-0059
USAMRAA
Grant ID: HT9425-24-C-0072
Disclaimer
This work is supported by the US Army Medical Research and Development Command under Contract No.W81XWH-21-C-0001, W81XWH-22-C-0093, HT9425-23-C-0059 and HT9425-24-C-0072. The views, opinions and/or findings contained in this report are those of the author(s) and should not be construed as an official Department of the Army or Navy position, policy or decision unless so designated by other documentation.
Abstract
This protocol describes flying vector storage.
Guidelines
OBJECTIVE
To outline steps for properly storing blood samples and nucleic acid samples purified from these samples.
SUMMARY/SCOPE
The overarching aim of the REDI-NET is to develop a collaborative laboratory network between domestic and international partnering institutions to address disease surveillance needs in order to effectively detect, predict and contain potentially emergent zoonosis. This SOP provides guidance on storage of blood samples and their purified nucleic acid to preserve their integrity for downstream nucleic acid extraction and/or sequencing library preparation
MAINTENANCE OF EQUIPMENT
Decontaminate a PCR workstation by keeping the UV light on for 00:15:00 .
Materials
EQUIPMENT AND MATERIALS
Note
NOTE: If product number is listed, please ensure use of this or equivalent product.
| A | B | C | |
| Equipment / Material | Description | Mfg / Product # | |
| –80°C freezer | For sample storage | Locally sourced | |
| Forceps | Clean, stainless | Locally sourced | |
| Ice | To maintain cold chain during sample handling | Locally sourced | |
| 96-Well Microfuge tube racks with cover | To hold microplates | Locally sourced | |
| KingFisher™ 96 KF microplate | To store the sample | ThermoFisher, 97002540 | |
| PCR Workstation | PCR cabinet with UV light | Locally sourced | |
| Clear Adhesive Film | To seal the KingFisher™ 96 KF microplate | ThermoFisher, 4306311 | |
| Adjustable micropipettes | To handle the samples | Locally sourced | |
| Multi-channel micropipettes | 8- or 12- channel; to handle the sample | Locally sourced | |
| Nuclease-free filter tips low-retention | To ensure appropriate sample handling | Locally sourced | |
| Nuclease free microfuge tubes | 1.5 mL | Locally sourced | |
| Saran wrap | Plastic wrap; to seal rack holding sample | Locally sourced | |
| Permanent markers | To label tubes and microplates | Locally sourced | |
| Data sheet | REDI-NET DCS T-3 Tick Storage | REDI-NET Data Portal |
Safety warnings
RISKS AND PERSONAL PROTECTION:
Gloves should be worn all the time when handling samples.
Ethics statement
The protocols.io team notes that research involving animals and humans, including tissues or samples collected directly from specimens or participants, must be conducted according to internationally-accepted standards and should always have prior approval from an Institutional Ethics Committee or Board.
STORAGE PROCEDURE FOR UNTREATED SAMPLE
Note
NOTES:
- Flying vector samples need to be kept on a cold chain all the time to prevent RNA degradation. The following procedure will apply only where -80ºC storage is feasible.
- If -80ºC storage is not possible, temporarily store the flying vector samples in a -20 ºC freezer and follow flying vector sample processing SOP (REDI-NET SOP FF-2 Flying Vector Processing) as soon as possible for total nucleic acid extraction. Subsequently, use a portion of the total nucleic acid and reverse- transcribe RNA into cDNA for -20 ºC storage. To do this, follow the initial steps of the flying vector sample testing SOP (REDI-NET SOP FF-4 Flying Vector Testing) till finishing the step of “Removal of RNA”.
Precool 96-well microfuge tube racks On ice .
Using preprinted adhesive labels or permanent markers, label 1.5 ml microfuge tubes with unique sample ID.
Transfer flying vector sample into the corresponding pre-labeled 1.5 mL microfuge tubes, close the cap and put it onto the microfuge tube rack (On ice ) sequentially.
Once the rack is full or all flying vector samples have been completed, label the rack with a unique rack ID.
Close the rack lid tightly, secure with clear Saran wrap and immediately transfer to -80 °C freezer.
Update the freezer inventory so samples can be tracked properly.
STORAGE PROCEDURE FOR TOTAL NUCLEIC ACID
Note
NOTES:
- The following procedure is to properly store total nucleic acid extracted from blood samples (including negative controls) using KingFisher nucleic acid purification system. The eluted total nucleic acid will be in either 96-well microplate (Flex model) or elution strip (Duo Prime model).
- Total nucleic acid samples need to be kept On ice all the time to minimize RNA degradation.
In the clean PCR workstation, carefully transfer the eluted total nucleic acid to a 96-well PCR microplate, make sure to keep samples in the exact same locations corresponding to the rack where the original blood samples were stored.
Note
IMPORTANT: Mark the “A1” position of the 96-well microplate to prevent any mistakes on plate orientation.
Cover the 96-well PCR microplate with adhesive film to prevent spill over or contamination.
Label the film with a unique plate ID.
Immediately transfer the 96-well PCR microplate to -80 °C freezer.
Update the freezer inventory so samples can be tracked properly.
Protocol references