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Sample | Processing number | Batch | Index |
|---|---|---|---|
1 | 1 | ||
2 | 1 | ||
3 | 2 | ||
4 | 2 | ||
5 | 3 | ||
6 | 3 | ||
7 | 3 | ||
8 | 3 | ||
9 | 3 | ||
10 | 3 | ||
11 | 3 | ||
12 | 3 | ||
13 | 3 | ||
14 | 3 | ||
15 | 3 | ||
16 | 3 | ||
17 | 3 | ||
18 | 3 | ||
19 | 3 | ||
20 | 3 | ||
21 | 4 | ||
22 | 4 | ||
23 | 4 | ||
24 | 4 | ||
25 | 4 | ||
26 | 4 | ||
27 | 4 | ||
28 | 4 | ||
29 | 4 | ||
30 | 4 | ||
31 | 4 | ||
32 | 4 | ||
33 | 4 | ||
34 | 4 | ||
35 | 4 | ||
36 | 4 |
1Cell Preparation
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- Thaw and stage capture and lysis materials
- Thaw one PIP tube per sample and RNase Inhibitor on ice
- Stage at RT Partitioning Reagent, Chemical Lysis Buffer 3
- Stage Combination 1.5 ml/0.5 ml tube stand, P200 and P1000 wide and standard
- Stage one 0.5 ml tube per sample
- Warm Cell Suspension Buffer to 37 C and cell suspension if frozen for 60-90 seconds
- decontaminate cell vial with 70% isopropyl alcohol
- thaw remaining ice at RT 30-60 sec
- Use a wide bore P1000 to gently mix and transfer to 15 ml conical tube
- Slowly add 9 ml warmed thawing media (>30s)
- Centrifuge cells at 200 xg for 5 minutes to pellet
- Aspirate as much supernatant as possible without disturbing pellet
- Add 1 ml prewarmed Cell Suspension Buffer, gently mix with wide bore P1000
- Place remaining Cell Suspension Buffer on ice
- Centrifuge at 200 x g for 3 min
- Aspirate as much supernatant as possible
- Use standard bore P100 to add 200-400 ul cold Cell Suspension Buffer and gently mix to resuspend 10-15 times
- Measure cell count
- Use wide bore P1000 to prepare 1,250 live cells/ul in Cell Suspension Buffer
- Measure cell count, repeat 9 and 10 until cell count reached
- place on ice
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