Nov 21, 2022
Biospecimen Aliquotting
- Clemens Scherzer1,2,
- Bradley Hyman3,2,
- Charles Jennings1,2
- 1Brigham and Women's Hospital;
- 2Harvard Medical School;
- 3Massachusetts General Hospital
Protocol Citation: Clemens Scherzer, Bradley Hyman, Charles Jennings 2022. Biospecimen Aliquotting. protocols.io https://dx.doi.org/10.17504/protocols.io.dm6gpwyz5lzp/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: February 18, 2021
Last Modified: May 31, 2024
Protocol Integer ID: 47413
Keywords: aliquot, aliquotting, bio, specimen, biospecimen, ASAPCRN
Abstract
This protocol explains the Standard Operating Protocol for aliquotting Bio-Specimens.
Guidelines
FREEZER STORAGE
Freezers are divided into 4 shelves, with 6 racks per shelf, and 24 boxes that can be held in each shelf. In total, 576 boxes, approximately 2,160 sample sets, can be stored in one -80°C freezer. The first three shelves are designated by visit number: Shelves A1-6 (top shelf) house samples from enrollment visits, shelves B1-6 (2nd shelf) house samples from the 1st year follow-up, and shelves C1-6 (3rd shelf) house samples from the 2nd year follow-up. Shelves D1-6 contain packed red blood cell tubes (PRBC), DNA, and RNA, extracted from blood as described in the protocols above. CSF is designated between two freezers in selected racks. Freezer storage and transactions of samples are recorded in the Freezerworks Inventory software.
Materials
MATERIALS:
- Freezer boxes (USA Scientific, Cat #9023-4981)
- 1.5 mL low-retention microcentrifuge tubes (Fisher Scientific, Cat #02-681-320)
- Low retention pipette tips
a. 1000 mL low-retention tips (Bio Plastic, Inc., Cat #3606SRS)
b. 200 uL low-retention tips (Molecular BioProducts, Cat #3932-05)
4. Freezerbonz labels (Fischer Scientific, Cat #22500521
Safety warnings
Please refer to Safety Data Sheets (SDS) for health and environmental hazards. Gain all required consent and experimental approvals before beginning any procedures.
1-2 days before aliquotting:
1-2 days before aliquotting:
Order dry ice the night before, at the latest, for delivery at 9 am.
Compile list of parent aliquots according to freezer position. Group aliquots from the same rack, column and box in the freezer.
Pull parent aliquots from separate locations and pool into one or more boxes. Place in one location in the freezer.
Print labels and label tubes for sub-aliquots.
In order to save time, scan barcodes of sub-aliquots into freezerworks.
Highlight parent aliquot to be sub-alquotted.
Click sub-aliquot.
Type in number of sub-aliquots, volumes (current and initial), freeze thaws, and amount to be deducted from parent aliquot.
Scan sub-aliquots into freezerworks.
Position sub-aliquots (position one sub-aliquot in parent aliquot’s original position).
Day of aliquotting:
Day of aliquotting:
2h
2h
Place parent aliquots On ice to thaw for about 02:00:00 .
2h
Sub-aliquotting parent aliquots:
Pipette samples in numerical order by sub-aliquot barcode label.
Any sample left over in the parent tube should be distributed evenly amongst the subaliquots.
Extra sub-aliquots can be created if remaining amount of sample in parent tube equals or exceeds desired volume.
Do not discard parent tube. Place in tube rack and set aside for scanning.
Do not discard extra, empty sub-aliquot tubes. Place in tube rack and set aside for scanning.
Place one new sub-aliquot in the original position previously held by the parent aliquot.
Place remaining sub-aliquots immediately into the -80 °C freezer in positions determined in Step 5.5.
Scan parent aliquot in “select by scanned field” function. Update parent aliquots by setting “current amount” to 0.
Scan extra, empty sub-aliquots in “select by scanned field” function. Delete aliquots entirely.
Add transactions in Freezerworks for each aliquot given to a collaborator outlining:
Date of aliquotting.
Type of transaction (i.e. “imparting of specimens”).
A note that describes to whom the aliquot is for, the volume, and type of aliquot.
(see “Adding transactions” protocol for additional information.)