Hello All, I’m hoping to get some help on pipetting sequences using Venus4.
We process a lot of plates by row where 96-well plate 1 has different reagents in Col1. These should be applied to 96-well plate B such that the reagent in plate 1, A1, will be used to fill Plate2 A1-12. Plate1,B1 will be used to fill Plate2 B1-12 etc. I’m hoping to use multi-dispense for all 12 dispenses.
The number of reagents is somewhat variable. For example it may only be 4 in plate 1, (A1,B1,C1,D1).
I can’t seem to figure out how to do this efficiently.
For a full plate, using the SmartStep aliquot I can get it to work in two pipette steps, where the first step takes A1,C1,E1, and G1 into rows A,C,E, and G of Plate2, then a second step that takes B1,D1,F1,H1 into rows B,D,E, and G of Plate 2.
However, this makes it difficult to edit based on the number of reagents in plate 1. Any help would be greatly appreciated!
Hi TDUB,
What edits are you trying to make specifically that isn’t working? Like are you trying to dynamically edit the destination sequences in plate 2 based on how many reagents you have in plate 1?
The step is intended to transfer 10 ul of well A1 in plate 1 to A1-12 of plate 2 and so on through H1. Ideally I could use a multidispense for this step to speed things along. Since the reagents in col1 of plate 1 are different in each row (A1 has different material than B1), I would like to use the same tip and channel across the entire row when aliquotting to plate2.
I had a hard time getting this done with a single step (it seems like it should be possible but I couldn’t figure it out). So I ended up with 2 steps. One for row for ACEG, and one for BDFH. Sometime I only have less than 8 reagents. So for example if I have reagents in A-E, then plate 2 should only get filled in rows A-through E. The way I have it now, it requires a manual adjustment of the sequences. I’m assuming I
Missing something, or maybe the aliquot step is not the right approach?
As long as the aliquot volume is the same throughout plate 2 and it’s in every set of 12 (12 wells per row), it is definitely possible to program it as single steps. Although smart steps does the calculation for you, it does have some limitation, notably channel variables (see below for more details).
https://forums.pylabrobot.org/t/channel-variables-for-smart-steps/1999?u=nat
I could imagine that if you have 7 reagents to work with but 8 channels activated in the smart step, the first 7 channels will aspirate in A1-G1 (plate 1) and then the the last channel will aspirate in A1, leading to a dispensing pattern that you don’t want.
I’m not currently near my instrument to provide further help (unless someone beats me to it) but, in the meantime, here’s an example of how to program a simple aliquot step using single steps.
https://forums.pylabrobot.org/t/liquid-following-stop-working-during-run/2713/12?u=nat
Also if there a reason for splitting the smart step into two steps for a full plate as opposed to doing it as one smart step (A-H)? Unless you are only working with 4 channels.