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What is the importance of emulsion-safe plastic consumables?

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.   

 

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Rainin pipette tips: updates for recent backorders

We are aware that there is currently limited availability of Rainin pipette tips. Rainin pipette tips have been extensively validated by 10x Genomics and are highly recommended for all single-cell assays. If Rainin tips are unavailable, any of the listed alternate pipette tips validated by 10x Genomics may be used.

 

Supplier Part Number
  Rainin
  • 20 µL, Catalog No. 30389226
  • 200 µL, Catalog No. 30389240
  • 1000 µL, Catalog No. 30389213
  Eppendorf*
  • 20 µL, Catalog No. 0030078535
  • 200 µL, Catalog No. 0030078551
  • 1000 µL, Catalog No. 0030078578
  Labcon
  • 20 µL, Catalog No. 4-1143-965-008
  • 200 µL, Catalog No. 4-1144-965-008
  • 1200 µL, Catalog No. 4-1145-965-008
  VWR**
  • 20 µL, Catalog No. 89495-374
  • 200 µL, Catalog No. 89495-378
  • 1000 µL, Catalog No. 89495-382
  Biotix
  • 20 µL, Catalog No. 63300931
  • 200 µL, Catalog No. 63300001
  • 1200 µL, Catalog No. 63300004

*Eppendorf pipette tips require Eppendorf pipettors. All other tips are compatible with Rainin pipettors.

**These VWR tips have been discontinued.

 

For steps downstream of the GEM-RT incubation step, it may be possible to use pipette tips from other vendors if Rainin tips or the alternate tips are in limited supply. We anticipate that any other pipette tips suitable for molecular biology assays would be compatible with downstream steps in 10x assays.

We have not tested alternative tips for the Visium workflows. However, it is low risk that application performance is impacted since no emulsion handling or microfluidics are involved.

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, Single Cell Multiome ATAC+GEX, Fixed RNA Profiling Gene Expression