AlanFung:LabNotes/Rolony/2013-11-14: Difference between revisions
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===Stripping=== | ===Stripping=== | ||
===Annealing=== | ===Annealing=== | ||
*Pre-heat padlock probes at 80C for 5 mins | |||
===Imaging=== | ===Imaging=== |
Revision as of 21:43, 14 November 2013
Optimum Probe# Test
- In this experiment we are trying to find the concentration of probes that works best for capturing rolony targets. From previous experiments we know that a concentration of 0.6uM with a volume of 100ul works.
- A test of 5 different concentrations will be perform and image analysis using PISA will be performed to count the number of target being captured
MALAT1 Padlock Probes Concentrations to test
5uM |
100nM |
10nM |
1nM |
0.1nM |
Protocol
- In this experiment we will be using the Mattek dish containing primary rolonies made on Oct 23rd dish #2
Reagents
80% Formamide in 2X SSC
MALAT1 padlock probes MALAT1 padlock probes detection
Apparatus
- warm up incubator to 50C for incubation
- warm up tube heater to 80C for pre-heating padlock probes
Stripping
Annealing
- Pre-heat padlock probes at 80C for 5 mins