Question: What is the importance of emulsion-safe plastic consumables?
Answer: Some plastics can interact with and destabilize GEMs. It is therefore critical to use validated emulsion-safe plastic consumables when handling GEMs. 10x Genomics has validated Eppendorf twin.tec® PCR plates and Rainin LTS Low retention pipette tips as GEM-compatible plastics. USA Scientific, Eppendorf, and Thermo Fisher PCR 8-tube strips have also been validated. Substituting these materials can adversely affect performance.
We have seen plastics that can negatively impact emulsions stability. These include brands from Biorad, Denville, and Axygen. The image below shows GEMs that were transferred to Denville branded tubes. The image was taken immediately after RT. Axygen tubes saw similar deterioration in emulsion stability after 24 hours, although nothing visually notable after RT or up to 12 hours.
Rainin pipette tips: updates for recent backorders
We are aware that there is currently limited availability of Rainin pipette tips. However, it is essential to use Rainin tips when handling GEMs. Other brands of pipette tips can destabilize GEMs, which can lead to a loss of single-cell behavior in the data.
In addition to the Rainin filter tips listed in our User Guides, we have also tested Rainin non-filter tips (part number 30389245) and found that these tips did not destabilize GEMs. Therefore, these non-filter tips can be used as an alternative tip if Rainin filter tips are not available.
For steps downstream of the GEM-RT incubation step, it may be possible to use pipette tips from other vendors if Rainin tips are in limited supply. At this time, we have not formally validated other brands of pipette tips for use with 10x assays and therefore cannot guarantee assay performance. However, we anticipate that any other pipette tips suitable for molecular biology assays would be compatible with downstream steps in 10x assays.
Therefore if you have limited access to Rainin tips, we would recommend saving/conserving your Rainin tips and using them only for steps involving handling GEMs. Tips from other vendors may be used for other steps in the assay.
Products: Single Cell Gene Expression, Single Cell Immune Profiling, Single Cell ATAC, Multiome