#503 closed enhancement (fixed)
Change pooling strategy
Reported by: | Nicklas Nordborg | Owned by: | Nicklas Nordborg |
---|---|---|---|
Priority: | major | Milestone: | Reggie v2.13 |
Component: | net.sf.basedb.reggie | Keywords: | |
Cc: |
Description
The current pooling protocol has a problem with libraries that have a low concentration. Part of the problem can be solved by looking at the entire pool as a group instead of only a single lib at a time when calculating how much EB to use.
The current target volume and concentration is 5µl and 2nM. Libraries with a concentration lower than 2nM cause the final pool concentration to decrease since we can't use more than 5µl. If we instead can use up to 10µl (all) for some libraries but decreasing the amount of EB used for the other libraries the total pool volume and concentration can be kept at optimal level in more cases.
Change History (9)
comment:1 by , 11 years ago
Status: | new → assigned |
---|
comment:2 by , 11 years ago
comment:3 by , 11 years ago
(In [2040]) References #503: Change pooling strategy
Better handling of the special case when the total EB volume is negative. This is probably an indication that the libraries are too bad to be sequenced. In the GUI a warning message is displayed and the pool summary is calculated as if EB volume is 0. Pool creation is still possible. The lab protocol wizard has also been fixed so that 0 is displayed for all EB volumes in this case.
comment:4 by , 11 years ago
comment:5 by , 11 years ago
comment:6 by , 11 years ago
comment:7 by , 11 years ago
(In [2053]) References #503: Change pooling strategy
Updates to the pooling lab protocol to make it clearer how EB should be mixed with libraries. When dynamic mixing strategy is used, no EB is displayed for individual libraries. Instead, the final EB is calculated and added to the pool after all libraries. The exception is high-concentration libraries that must be mixed to double volume before adding them to the pool. This is always displayed as a special "Remark" and not in the volume/eb columns.
comment:8 by , 11 years ago
Resolution: | → fixed |
---|---|
Status: | assigned → closed |
(In [2027]) References #503: Change pooling strategy
The new implementation works like this:
There is currently not enough checking for extreme cases, eg. if there are many libraries with low concentration the final EB volume may be negative.
NOTE! Since the new implementation is very different from the old, case is needed when installing this update. Pools that have been created with the old Reggie wizard should be completed in the lab and registered before installing this update. Otherwise the lab protocols may show incorrect mixing values.
The incorrect calculations for 'dil' (see #507) items have been fixed.