Nov 24, 2025
Version 1
Karyotyping Protocol V.1
- Kamal Gill1
- 1Clelland Lab
Protocol Citation: Kamal Gill 2025. Karyotyping Protocol. protocols.io https://dx.doi.org/10.17504/protocols.io.5qpvodmy7g4o/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: November 24, 2025
Last Modified: November 24, 2025
Protocol Integer ID: 233364
Keywords: karyotyping protocol clelland lab, protocol clelland lab internal protocol, protocol, internal protocol
Abstract
Clelland Lab internal protocol for karyotyping.
Materials
Items you need to bring/find
1. Shipping cardboard box with styrofoam box inside. It needs to fit a T-25 standing
2. Small ziplock bags (that can fit a T-25)
3. FedEx slip + FedEx Slip pouch (it has an adhesive side to attach to the box)
4. Paper towels, bubble wrap or similar product (can find by asking other labs nicely)
5. Parafilm
6. Shipping Scale (located on the 1st floor. Take the service elevator down and it should be in that corridor)
7. Shipping tape (should be on the first floor next to the shipping scale)
Media/Reagent
1. Essential 8™ Medium (Thermo A1517001)
2. Fetal Bovine Serum (FBS) (Cytiva SH30071.03HI) (if you don't have this, that is ok)
Items at Gladstone Stem Cell Core
1. T-25 with sealed cap (should be in SCC, if not that is also OK)
2. Ethanol
3. 50mL Conical Tube
Instructions
Make sure the T-25 is 40-60% confluent
Warm 100mL E8 + 15% FBS (85 mL of E8 with 15mL FBS) to 37°C.
I usually use the bead bath
Let this incubate for 30 minutes, but you can cut down this step if you don't have time
Locate a cardboard shipping box that is small and has a styrofoam box within it.
You cannot just use a styrofoam box
Clean the box with ethanol and remove old shipping information from the box
In the stem cell core, find a T-25 with sealed caps (it looks just like a regular T-25, but the cap has no filter)
If you can a T-25 with a sealed cap
You only need the caps of the sealed T-25. You will toss the bodies.
You will swap caps with this sealed T-25 after filling your sample with media
If you cannot find a T-25 with a sealed cap
You can use extra parafilm
After the media is warm, aspirate spent media from T-25 in a BSC
Hold the T-25 so the cap is facing upwards, then slowly fill the T-25 with warm E8+ 15% FBS.
If you have a sealed cap, fill up the T-25 until there is little room at the cap. Toss the excess plasticware
If you do not have a sealed cap, leave a centimeter - 1.5 centimeters of space.
NOTE: Avoid aiming the stream at cells! That can cause cells to lift.
Place excess media in a 50mL conical tube. This will be shipped as well.
Prepare items for shipping
Take the T-25 and conical tube to a bench area.
Use parafilm to fully seal the tops of the T25 and Conical tube. Be extra conscious of sealing when using the non-sealed T-25 cap.
Place the items in small ziplock bags
Submit a Characterization request form via WiCell’s website
Place items in the box
Layer the shipping box with paper towels
Place T-25 and the excess media
Add bubble wrap or similar product to prevent items from shifting during shipping
Attach ship form from WiCell’s website
Seal the box
Ship box: fill out the FedEx slip
Enter WiCell’s address
Select FedEx priority overnight
Other packaging
Select dangerous good, but shipping declaration not required
Determine the weight of the box using a scale
Charge sender
You are done! You can leave the box at the FedEx pick up location on the 1st floor at Gladstone!