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Multiplication

Subculturing (or recycling) is carried out at the end of the storage cycle, when accessions show obvious signs of deterioration and/or when stock becomes low and there is a need for multiplication or safety duplication.

tip

It is prudent to maintain a few viable and healthy cultures of the previous subculture cycle as “spare materials” until the newly subcultured set is healthy and growing.

Subculturing​

  1. Set aside the germplasm you will subculture
  2. Open In vitro > Multiplication and scan the barcode of the material to be subcultured
    • You can use multiple plants of the same inventory by scanning the multiple tubes
  3. Select the number of new inventories (new separate lines) to be registered
    • Use 1 to track the new batch of material under the same identity and barcode

Registering subcultured material

  1. Select the maintenance policy and enter the quantity of each new inventory
    • Culture media, type of container, etc. can also be specified

Enter details for each line

  1. Click Preview and Save
    • Barcode labels for the new inventories will be automatically prepared
note

Depending on the Auto-deducted flag, the quantity of the source material may not be automatically reduced.

Tracking parentage​

These tools automatically maintain the relationships between the source and subcultured germplasm. The Inventory tree of the newly registered line GBK 120.2 1 IV from the example above shows that:

  • it was sourced from "GBK 120 2 IV" which came from "T2025 20 IV",
  • its generation number (3) was automatically incremented from source's (2),
  • and its propagation date was automatically recorded (not shown)

Germplasm relationships

Naming convention​

The inventory prefix and sequence number of the source inventory are concatenated and used as the prefix for the new generation. The next number in the sequence is automatically assigned. In the example above we multiplied an inventory named GBK 120 2 IV and the subcultured inventory is called GBK 120.2 1 IV:

  • The inventory prefix GKB 120.2 comes from the prefix (GBK 120) and sequence number (2) of the source inventory.
  • The number 1 was automatically assigned.
  • The IV represents the inventory type "In vitro"

When GBK 120.2 1 IV is subcultured, the new lines will be called:

  • GBK 120.2.1 1 IV
  • GBK 120.2.1 2 IV
  • GBK 120.2.1 3 IV, etc.

Subculturing GBK 120.2.1 3 IV results in:

  • GBK 120.2.1.3 1 IV
  • GBK 120.2.1.3 2 IV and so on.

Rejuvenation​

For cultures requiring rejuvenation (transfer of accessions to the greenhouse and field, followed by re-initiation into tissue culture) please follow the distribution approach:

  1. Create a new distribution request
  2. Add the materials to be rejuvenated by scanning their barcodes
  3. Create withdrawn inventories to maintain the link between the source germplasm and the material in the greenhouse
    • Print labels for the withdrawn inventories
  4. Complete the distribution process and rejuvenate the material according to your protocols
  5. Re-introduce it to in vitro after successful rejuvenation