text stringlengths 0 4.18k |
|---|
Tab 2 Text |
Tab 3 Text |
Tab 4 Text |
Tab 5 Text |
Tab 6 Text |
## Index Hopping Mitigation |
Index hopping can impact pooled samples sequenced on Illumina sequencing platforms that utilize patterned flow cells and exclusion amplification chemistry. To minimize index hopping, follow the guidelines listed below. |
- l Remove adaptors during cleanup steps. |
- l Ensure no leftover primers and/or adaptors are present when performing Post-Library Construction QC. |
- l Store each library individually at 4°C for up to 72 h or at -20°C for longterm storage. DO NOT pool libraries during storage. |
- l Pool libraries prior to sequencing. An additional 1.0X SPRI may be performed for the pooled libraries to remove any free adaptors before sequencing. |
- l Hopped indices can be computationally removed from the data generated from single cell dual index libraries. |
Tab 1 Text |
Tab 2 Text |
Tab 3 Text |
Tab 4 Text |
Tab 5 Text |
Tab 6 Text |
<!-- image --> |
## Step 1: |
## GEM Generation and Barcoding |
| 1.0 Get Started | 30 | |
|------------------------------------------|------| |
| 1.1 Prepare Master Mix | 31 | |
| Assemble GEM-X Chip | 32 | |
| 1.2 Load GEM-X Chip | 36 | |
| 1.3 Run the Chromium X Series Instrument | 38 | |
| 1.4 Transfer GEMs | 39 | |
1 |
1.5 GEM-RT Incubation |
40 |
## 1.0 Get Started |
<!-- image --> |
Firmware Version v2.0.0 or higher is required for Chromium X/iX and v2.2.0 or higher is required for Chromium Xo instrument. |
| Item | 10x PN | Preparation & Handling ... |
|---------------------------------|------------------------------------------------|--------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------... |
| Equilibrate to Room Temperature | Equilibrate to Room Temperature | ... |
| □ | GEM-X Single Cell 3' Gel Bead v4 2001128 | Equilibrate to room temperature 30 min before loading the chip. ... |
| □ | RT Reagent E 2001106 | Vortex, verify no precipitate, centrifuge briefly. If a precipitate is observed, warm the tube with hands until the precipitate dissolves. Vortex and centrifuge ... |
| □ | Template Switch Oligo B 2001027 | Centrifuge briefly, resuspend in 65 μ l Low TE Buffer. Vortex 15 sec at maximum speed, centrifuge briefly. Resuspended solution can be used immediately. After resuspension, store at -80°C . Thaw at room temperature for ≥ 30 minutes in ... |
| □ | Reducing Agent B 2000087 | Vortex, verify no precipitate, centrifuge briefly. ... |
| Place on Ice | Place on Ice | ... |
| □ | Cell Suspension - | ... |
| □ | RT Enzyme E 2001105/ 2001146 | Centrifuge briefly before adding to the mix. ... |
| □ | 1X PBS - | ... |
| Obtain | Obtain | ... |
| □ | Partitioning Oil B 2001213 | - ... |
| □ | GEM-X 3' Chip Verify name and PN 2001097 | See Tips & Best Practices. ... |
| □ | X/iX Chip Gasket 3000656 | See Tips & Best Practices. ... |
| □ | Chromium X/iX or Chromium X Series Chip Holder | 3000598 See Tips & Best Practices. ... |
| □ | 10x Vortex Adapter 330002 | See Tips & Best Practices. ... |
| □ | 50% glycerol solution If using < 8 reactions - | See Tips & Best Practices. ... |
Tab 1 Text |
Tab 2 Text |
Tab 3 Text |
Tab 4 Text |
Tab 5 Text |
Tab 6 Text |
## 1.1 Prepare Master Mix |
- a. Prepare Master Mix on ice. Pipette mix 15x and centrifuge briefly. |
<!-- image --> |
<!-- image --> |
<!-- image --> |
<!-- image --> |
Subsets and Splits
No community queries yet
The top public SQL queries from the community will appear here once available.