Question: Can I sequence V(D)J-enriched and 5' Gene Expression libraries on the same lane?
Answer: Yes. V(D)J-enriched libraries and 5’ Gene Expression libraries can be pooled for sequencing, but note the depth requirements and sequencing configurations are different. We recommend 5,000 reads pairs/targeted cell for V(D)J-enriched libraries and a minimum of 20,000 read pairs/cell for 5’ Gene Expression libraries, and therefore recommend adjusting the pooling ratio accordingly. (For example, if the two library types have the same number of cells targeted and the same library concentration, 4-fold more 5' Gene Expression Library should be mixed with the V(D)J-enriched Library to obtain 4x more reads per cell.)
Our recommended parameters for sequencing V(D)J and 5' Gene Expression libraries together is as follows:
- v1 and v1.1: Read 1: 26 cycles, i7 Index: 8 cycles, Read 2: 91 cycles
- v2 (dual index): Read 1: 26 cycles, i7 Index: 10 cycles, i5 Index: 10 cycles, Read 2: 90 cycles
Please see our page on sequencing requirements for V(D)J libraries for more information on sequencing options.
Products: Single Cell Immune Profiling