AlanFung:LabNotes/Capturing/2014-1-23: Difference between revisions
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*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. | |||
==ssDNA Qubit to quantify probe and template== | ==ssDNA Qubit to quantify probe and template== | ||
==Probe calculations ppMALAT1== | ==Probe calculations ppMALAT1== |
Revision as of 20:23, 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 | 0.99 | 100 | 30 | 29.01 |
MALAT1 Template | 1485 | 1.35 | 100 | 20 | 18.65 |
ppillumina_155 | 3267 | 0.92 | 100 | 30 | 29.08 |
10BP | 1000 | 0.10 | 100 | 1 | 99 |
- 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.