Protocol
A correction was made to Transcriptome Analysis of Single Cells. There were errors in the volumes used in the aRNA Amplification section. The volumes were changed from:
- 6.2.1 7.5 μL 5x Second strand buffer
- 6.2.4 Add 29 μL DEPC water
2 μL glycogen (5 mg/mL) 1/10 volume (3 μL) 3M Sodium acetate
2.5 volumes (~250 μL) cold 100% ethanol - 6.2.7 Remove the supernatant and air dry for 15-20 minutes.
- 6.3.4.2 Add 50 μL DEPC water
2 μL glycogen (20 mg/mL)
0.6 volumes (37.2 μL) 5M Ammonium acetate
2.5 volumes (~180 μL) cold ethanol - 6.3.4.7 Remove supernatant and airy dry 15-20 minutes.
- 6.3.5.1 Add 50 μL DEPC water
2 μL glycogen (20 mg/mL)
0.6 volumes (37.2 μL) 5M Ammonium acetate
1 volume (98 μL) phenol:chlorofom:isoamyl alcohol 25:24:1 (equilibrated to pH 7.8-8.0) - 6.3.5.7 Remove supernatant and air dry 15-20 minutes.
to
- 6.2.1 5.6 μL 5x Second strand buffer
- 6.2.4 Add 40 μL DEPC water
1 μL glycogen (5 mg/mL) 1/10 volume (5 μL) 3M Sodium acetate
2.5 volumes (~125 μL) cold 100% ethanol - 6.2.7 Remove the supernatant and air dry
for 15-20 minutes. - 6.3.4.2
Add 50 μL DEPC water
2 μL glycogen (5 mg/mL)
0.1 volumes (10 μL) 5M Ammonium acetate
2.5 volumes (~275 μL) cold ethanol - 6.3.4.7 Remove supernatant and air dry
15-20 minutes. - 6.3.5.1 Add 90 μL DEPC water
2 μL glycogen (5 mg/mL)
0.1 volumes (10 μL) 5M Ammonium acetate
1 volume (100 μL) phenol:chlorofom:isoamyl alcohol 25:24:1 (equilibrated to pH 7.8-8.0) - 6.3.5.7 Remove supernatant and air dry
15-20 minutes.
