AlanFung:LabNotes/Capturing/2014-1-23: Difference between revisions
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==ssDNA Qubit to quantify probe and template== | ==ssDNA Qubit to quantify probe and template== | ||
*ppMALAT1 (4.78ng/ul) /(0.4/120) = 1434ng/ul | *ppMALAT1 (4.78ng/ul) /(0.4/120) = 1434ng/ul vs 6072ng/ul (calculated based on stock concentration) | ||
*MALAT1 template (15.54ng/ul)/(0.4/30) = 1165.5ng/ul | *MALAT1 template (15.54ng/ul)/(0.4/30) = 1165.5ng/ul vs | ||
*ppillumina_155 (7.02ng/ul)/(0.4/65) = 1140.75ng/ul | *ppillumina_155 (7.02ng/ul)/(0.4/65) = 1140.75ng/ul vs | ||
==Probe calculations ppMALAT1== | ==Probe calculations ppMALAT1== |
Revision as of 22:53, 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, 200uM=6072ng/ul)
- MALAT1 Template (45nt, 100uM=1485ng/ul)
- ppillumina_155 (99nt, 100uM=3267ng/ul)
Sample | ng/ul | ul | ng/ul | ul | H2O |
ppMALAT1 | 6072 | 0.40 | 20 | 120 | 119.60 |
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.
File:ZhangLab 2 2014-01-23 13hr 49min.jpg
ssDNA Qubit to quantify probe and template
- ppMALAT1 (4.78ng/ul) /(0.4/120) = 1434ng/ul vs 6072ng/ul (calculated based on stock concentration)
- MALAT1 template (15.54ng/ul)/(0.4/30) = 1165.5ng/ul vs
- ppillumina_155 (7.02ng/ul)/(0.4/65) = 1140.75ng/ul vs