AlanFung:LabNotes/Capturing/2014-1-23: Difference between revisions
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| height="13" | ppMALAT1 | | height="13" | ppMALAT1 | ||
| align="center" | 3036 | | align="center" | 3036 | ||
|style="background-color:#EBF1DE" align="center" | 0. | |style="background-color:#EBF1DE" align="center" | 0.40 | ||
| align="center" | | | align="center" | 20 | ||
| align="center" | | | align="center" | 60 | ||
|style="background-color:#EBF1DE" align="center" | | |style="background-color:#EBF1DE" align="center" | 59.60 | ||
|- align="center" | |- align="center" | ||
| height="13" | MALAT1 Template | | height="13" | MALAT1 Template | ||
| align="center" | 1485 | | align="center" | 1485 | ||
|style="background-color:#EBF1DE" align="center" | | |style="background-color:#EBF1DE" align="center" | 0.40 | ||
| align="center" | 20 | | align="center" | 20 | ||
|style="background-color:#EBF1DE" align="center" | | | align="center" | 30 | ||
|style="background-color:#EBF1DE" align="center" | 29.6 | |||
|- align="center" | |- align="center" | ||
| height="13" | ppillumina_155 | | height="13" | ppillumina_155 | ||
| align="center" | 3267 | | align="center" | 3267 | ||
|style="background-color:#EBF1DE" align="center" | 0. | |style="background-color:#EBF1DE" align="center" | 0.40 | ||
| align="center" | | | align="center" | 20 | ||
| align="center" | | | align="center" | 65 | ||
|style="background-color:#EBF1DE" align="center" | | |style="background-color:#EBF1DE" align="center" | 64.6 | ||
|- align="center" | |- align="center" | ||
| height="13" | 10BP | | height="13" | 10BP | ||
| align="center" | 1000 | | align="center" | 1000 | ||
|style="background-color:#EBF1DE" align="center" | 0. | |style="background-color:#EBF1DE" align="center" | 0.40 | ||
| align="center" | | | align="center" | 20 | ||
| align="center" | | | align="center" | 20 | ||
|style="background-color:#EBF1DE" align="center" | | |style="background-color:#EBF1DE" align="center" | 19.6 | ||
|} | |} |
Revision as of 20:26, 23 January 2014
Capturing ppMALAT1 template w ppMALT1 ordered form IDT
TBU GEL to verify probe template size
- Aim for 100ng/band
- 10bp Ladder
- ppMALAT1 (92nt, 100uM=3036ng/ul)
- MALAT1 Template (45nt, 100uM=1485ng/ul)
- ppillumina_155 (99nt, 100uM=3267ng/ul)
Sample | ng/ul | ul | ng/ul | ul | H2O |
ppMALAT1 | 3036 | 0.40 | 20 | 60 | 59.60 |
MALAT1 Template | 1485 | 0.40 | 20 | 30 | 29.6 |
ppillumina_155 | 3267 | 0.40 | 20 | 65 | 64.6 |
10BP | 1000 | 0.40 | 20 | 20 | 19.6 |
- Pre-run the gel for 30 min at 200V
- Dilute sample to 20ng/uL
- Denature samples and ladder at 75C for 7 min, then quickly transfer to cooling rack, wait for 5 mins
- Flush the wells very well with P1000 & P200 pipette to remove all urea.
- NOTE: DO NOT TOUCH THE TUBE WITH YOUR HANDS IN THE FOLLOWING STEP, GRAB THE CAP
- Load the samples into the wells, run at 200V for 30 min.
- Stain gel with 3uL SYBR Gold in ~100 mL of TBE in clean tray.
- Place tray on an orbital shaker for ~5 min.