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-   -   Single-cell libraries miniaturization (http://seqanswers.com/forums/showthread.php?t=80328)

sbarberan 01-26-2018 11:16 AM

Single-cell libraries miniaturization
 
Any advice from single-cell experts about the best liquid handler for miniaturization of libraries?

We are considering:
Mantis (Formulatrix)
Mosquito (ttplabtech)

Any comments about either instrument?

Thanks in advance!
Sergio

Simone78 02-08-2018 10:13 AM

It depends which protocol you want to use.
If you buy a Chromium or do Seq-well/Drop-seq/InDrop/Nadia system, etc etc most likely you donīt need any robot since the library prep will be done in 1 tube and youīll never have such a high throughput. Itīs very likely you run out of money first because of the sequencing!:)

If you plan to do FACS sorting and use a plate-based method (Smart-seq2, MARS-seq, etc) you can choose among several nanodispensers:
- Mantis: cheaper but not sure it is really high throughput. It might take long to fill up many plates. We donīt have one, I just saw it in action, so take my comment with a grain of salt.
- Nanodrop (Innovadyne): amazing instrument. Non-contact dispenser, no tips, pretty fast: 30 sec to fill a 384w plate, can do up to 2 plates at the time. Very robust, lower limit pipetting 80-100 nL but it can go down to 50 nl (some modifications required). Uses 96, 384 and, with some tweaking, 1536 plates. We have it and we love it! No humidity chamber, though. Dispensing mineral oil is NOT recommended.
- Mosquito: we just got one for a trial. Slower than Nanodrop but cheaper (less than half the price), uses quite SOME tips but can dispense any solution due to the positive displacement pipetting system. Nanodrop, on the other hand, is in trouble with solution containing >10-15% glycerol. Can have humidity chamber, which is an advantage if you want to miniaturize your reactions. Lower limit pipetting is 25 nL (HTS) or 500 nL (HV)
- iCELL8. Uses the same type of dispenser as the Nanodrop but is much more precise and can dispense into 5184 well chips. Lower limit pipetting is 35 nl (theoretical) or 50 nl (real world). However, it uses special chips and requires specific kits (not exactly cheap...), has a higher price tag and you would need to dispense cells by limiting dilutions (= Poisson), which doesnīt make it very competitive, at the moment.

there are others but this is good enough for a start!:)
Best,
Simone

sbarberan 02-08-2018 02:10 PM

Thanks a lot Simone!

We want to use FACS sorting, and we are only looking for an instrument to reliably prepare 1/2 or 1/4 size libraries to reduce cost.

Cheers,
sergio

sbarberan 02-08-2018 02:46 PM

I also was wondering if there are any users with experience with single-cell printing (http://www.cytena.com/SCP.html) and how this could compare to FACS sorting. Looks to me that cells undergo less manipulation/stress with the printer than with FACS sorting, but I have not used the instrument myself. Any single-cell printer users?

Cheers,
sergio


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