Ligation sequencing V14 — single-cell transcriptomics with 3' cDNA prepared using 10X Genomics on PromethION (SQK-LSK114) (SST_9198_v114_revM_25Feb2025)

DateVersionLanguageDeviceChanges
4/9/2025SST_9198_v114_revM_25Feb2025EnglishPromethION
25th February 2025 In "Overview of the protocol" and "Equipment and consumables", compatibility information has been corrected to say "This protocol is intended for use with the 10X Genomics GEM-X Single Cell 3' Kit (V4), Next-GEM Single Cell 3' Kit (V3.1), or the Visium Spatial Gene Expression Kit (V1)."
1/29/2025SST_9198_v114_revK_29Jan2025EnglishPromethION
29th January 2025 In "Post pull-down PCR", the quantification step has been changed to say "Quantify 1 µl of eluted sample using a Qubit fluorometer - recovery aim >100 ng total.".
11/13/2024SST_9198_v114_revJ_13Nov2024EnglishPromethION
November 2024 - Updated concentration typos in step 3 of the "Pull-down" section of the protocol: Transfer 3.5 ml of 2X wash/bind buffer (__10 mM Tris-HCl pH 7.5, 2 M NaCl, 1 mM EDTA__) to a clean 15 ml Falcon tube.
11/13/2024SST_9198_v114_revI_06Dec2023EnglishPromethION
November 2024 - Updated concentration typos in step 3 of the "Pull-down" section of the protocol: Transfer 3.5 ml of 2X wash/bind buffer (__10 mM Tris-HCl pH 7.5, 2 M NaCl, 1 mM EDTA__) to a clean 15 ml Falcon tube.
11/6/2024SST_9198_v114_revI_06Dec2023EnglishPromethION
November 2024 - Formatting fix to custom-ordered oligo sequences table in equipment and consumables to prevent some of the sequence being cut off in the PDF generation.
8/21/2024SST_9198_v114_revH_06Dec2023EnglishPromethION
21st August 2024 - Added warning to the "Post-pull-down PCR" step to ensure users DO NOT allow the amplicon-bead conjugate to pellet before incubating. - NEB reagent recommendations for purchase and use updated (Salt T4 ligase in place of Quick Ligation Module). - Updated the Adapter ligation step with new recommended method.
August 21st, 2024SST_9198_v114_revG_31Jul2024English- Download 31 July 2024 - Changed reagent order addition in Library preparation/Adapter ligation and clean-up; Ligation Adapter (LA) added before Ligation Buffer (LB) and NEBNext Quick T4 Ligase 30 July 2024 - Updated wording in protocol introduction and in equipment and consumables: "This protocol is compatible and fully supported with 10X Genomics Next GEM Single Cell 3' Kit (V3.1) and the Visium Spatial Gene Expression Kit (V1). Other versions of the kits are not supported."
July 31st, 2024SST_9198_v114_revF_06Dec2023English- Download 30 July 2024 - Updated wording in protocol introduction and in equipment and consumables: "This protocol is compatible and fully supported with 10X Genomics Next GEM Single Cell 3' Kit (V3.1) and the Visium Spatial Gene Expression Kit (V1). Other versions of the kits are not supported."
July 30th, 2024SST_9198_v114_revE_06Dec2023English- Download 07 May 2024 - Updated library loading recommendation at the end of "Adapter ligation and clean-up" to use 50–100 fmol going into the flow cell.
May 14th, 2024SST_v9198_v114_revE_06Dec2023English- Download 07 May 2024 - Updated library loading recommendation at the end of "Adapter ligation and clean-up" to use 50–100 fmol going into the flow cell.
April 12th, 2024SST_v9198_v114_revB_06Dec2023English- Download February 2024 - Udated protocol to be specific to 3' cDNA (New 5' cDNA specific 10X protocol now also available on the community).
December 6th, 2023SST_v9198_v114_revA_06Dec2023English- Download -