Updating to MeLuDI 1.3.2

Updating to MeLuDI 1.3.2 might require some manual changes for items that already exist. This is due to the following decision (See ticket:802 for more information):

• Input of all lot numbers of interest is now performed in the first DNA/RNA extraction wizard, the "Lab tracking protocol for FFPE extraction" wizard, and the entered lot numbers are stored with the created start item list. Other DNA/RNA extraction wizards will obtain the default lot numbers from the input start item list.

Consequences for older start item lists:

• Previously, the lot number for the "FFPE DNA/RNA kit" entered in the first extraction wizard was stored as annotation for the created start item list, while all other lot numbers were entered in the following DNA/RNA registration wizard and stored as annotations for the created DNA/RNA extract items. Since individual entries for extracts were allowed, and still are, it was found better to store default lot numbers as annotations to the start item list. In order for the updated "Lab tracking protocol/report copy" wizard to be able to display all lot numbers for start item lists created prior to MeLuDI v1.3.2, the latter start lists need to be updated.

A new wizard, "Update processed start lists with lot numbers from extracts", has been added to the "Special conversion tools" sub-section of the "Admin" section. It allows a number of processed start item lists to be selected from a menu (normally all start item lists are selected, if the conversion is performed directly after installing MeLuDI v1.3.2). For each selected start item list, new lot number annotations of the latter will be set to values of the corresponding annotations for the first DNA extract, or if no such extract exists, the first RNA extract (MeLuDI items are used if existing, otherwise extra material items). The new extraction operator annotation will not be set, as it was not defined for extracts prior to MeLuDI v1.3.2. The conversion tool is only needed once, after update to MeLuDI v1.3.2, and can therefore be removed in later versions.

Older unprocessed start item lists do not state a problem, although they lack values for most lot numbers, since these will be added in the "DNA/RNA registration/quantification" wizard.

Follow this procedure:

1. Preparations for transferring lot numbers from first DNA extract to a processed start item list created prior to MeLuDI v1.3.2.

a. Update to MeLuDI v1.3.2

Among other things, annotation types for all lot numbers are installed for item lists.

Important!

1. MeLuDI v1.3.2 requires at least version 3.5.0 of BASE. If needed, update BASE before installing MeLuDI v1.3.2.
2. After installation of MeLuDI v1.3.2, run the installation wizard.

b. Check what start plates contain information on extra material, and collect the information

Select BASE menu option Biomaterial LIMS -> Bioplates to display a list of bioplates. Display values for name, BatchNumberExtraQiacubeItems [A], and ExtraQiacubeItemNamesListString [A]. Filter name for "LP%" to display start plates, and annotation BatchNumberExtraQiacubeItems [A] for ">0" to show start plates with data for extra material. Note values of names for extra material in annotation ExtraQiacubeItemNamesListString [A]. If not present here, check for extra material names in paper copies of lab tracking protocol for Allprep isolation. Make a list outside the BASE/MeLuDI software of the extra material names for each start plate, or, if no names are stored, the number of extra items.

c. Check if any extra material names need to be modified

If necessary, modify names for extra material according to the following:

1. If more than one extra material item has the same name, modify some of them.
2. If some extra material names start with "ML", modify them.
3. If some extra material core names contain special characters dot ".", comma, semicolon, or space, modify them.
4. If some extra material core names consist entirely of digits, modify them.
5. Construct names of extra material for start plates, where no names had been stored.
6. All previously used extra material were of type specimen, since they were QIAcube items. Therefore add a suffix "." + number to the core name. Start with number "1" for the first specimen having a specific core name, and increment the number for each new specimen with the same core name.

The result is a list of extra material specimen names for the start plates, where the former can be used to register extra material in MeLuDI v. 1.3.1

d. If needed, modify the start plate data to include the updated extra material names

Select BASE menu option Biomaterial LIMS -> Bioplates to display a list of bioplates. Display values for name, BatchNumberExtraQiacubeItems [A], and ExtraQiacubeItemNamesListString [A]. Filter name for "LP%" to display start plates, and annotation BatchNumberExtraQiacubeItems [A] for ">0" to show start plates with data for extra material. For each start plate:

1. Click on the start plate name, to display info for the start plate.
2. Select tab "Annotations". Under primary annotations, for "View" select radio button "All annotations".
3. For annotation ExtraQiacubeItemNamesListString, click the "Edit" icon at the right end of the value field, to open an edit dialog window. In the edit dialog, enter new data or modify existing to get a comma-separated line of the extra material names for the start plate, that were constructed in the previous section. Make sure to use the specimen names ending in "." plus a number. Click the "Save" button in the edit dialog, to save the new values and close the dialog window.

e. Register (optionally modified) extra material names

In section "Case registration" of the MeLuDI v.1.3.1 software, select the "Register new extra material" wizard. Enter data for the extra material specimens, whose names are included in the start plate annotation ExtraQiacubeItemNamesListString. Note that the core name without suffix should be entered in this wizard, since the suffix is added later, based on the chosen item type (here equal to specimen). Register the extra material. The wizard will now create the extra material specimens and child DNA and RNA extracts.

2. Create start item lists

a. Create start item list from old start DNA plate

In section "Server administrator wizards" of the MeLuDI v.1.3.1 software, select the "Create start lists from start DNA plates" wizard.

1. Select the start plates you want to create start item lists for (normally all the plates).
2. Click "Register" to create the start lists.

The following assumptions must be met for success:

1. The start plate contains a list of all related MeLuDI item names as annotation.
2. The start plate contains a list of all related extra material names as annotation.
3. All related specimens and extracts have already been created with the specified names.

For each start plate, a start item list will be created, with the following properties:

1. The start list name is that of the start plate with prefix "LP" exchanged for "SL".
2. All relevant start plate annotations are copied to corresponding start list annotations.
3. Items named in start plate annotations are added as start list members (DNA child extracts represent specimens).
4. An optional start plate creation date is stored in start list "ListExtractionDate" annotation.

The created start item lists can now be inspected using the "Lab tracking protocol/report copy" wizard.

Storage of the start plate creation date in an item list annotation was chosen, as creation events are not defined for item lists. Since a set creation date for a start plate in MeLuDI v1.2.* indicates that the plate has been processed, i.e. that concentration and delta-Ct values have been registered for MeLuDI DNA/RNA, the created start list should also be marked as processed, by setting value of Boolean annotation "SamplePrepListIsProcessed" to true, if the start list does not contain any extra material. However, this should not be done for start lists with extra material, since concentration and delta-Ct values were only registered for MeLuDI items in v1.2.*, but not for extra material. Setting the value of annotation "SamplePrepListIsProcessed" to true, would prevent the start list from being opened in the "DNA/RNA registration/quantification" wizard, in order to register concentrations and delta-Ct values for the extra material.

3. (Optional) Register quantification/QC values for extra material items

If desired, concentration and delta-Ct values can now be registered for the new extra material items.

a. Store kit lot numbers for each start item list in a file

The "DNA/RNA registration/quantification" wizard will display previously registered values for MeLuDI extracts in the input form as a practical side effect, so these values need not be re-entered. Unfortunately, the latter is not true for kit lot numbers. The following procedure is therefore recommended to register additional values for an item list:

1. Open each start item list with extra material in the "Lab tracking protocol/report copy" wizard.
2. In step 2, after existing kit lot number values have been loaded, click button to download values to a reagent lot number file. Cancel this wizard when ready.

b. Register concentration and delta-Ct values for extra material items

1. Open each start item list with extra material in the "DNA/RNA registration/quantification" wizard.
2. In step 2, select to import kit lot numbers from the file saved previously.
3. In step 3, enter the missing concentration and delta-Ct values for extra material, and complete the registration.

4. After controlling the created start item lists, delete or rename the old start DNA plate items

After controlling the created start lists, the start plates may be deleted. An alternative is to rename the plates by adding a new prefix (e.g. "OLD_"), so the names do not start with "LP". The start plates will reappear in the library preparation wizards, when they are to be filled with samples (though not necessarily the same samples as were previously related to it via annotations).