User:Pranav Rathi/Notebook/OT/2011/01/10/DNA tethering experiments
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I attempt tethering again today with new beads and new anti-dig.
Ant-dig
Dr. Koch and I prepared new anti-dig: 1X concentration
- PBS [1]180μl + aliquots 20μl = [MIX] = Anti-dig 200μl.
PBS was used from the glass door fridge and aliquot from super cold gray fridge. I used two different beads:
- 1 μm new beads (looks good in microscope), 20μl from the batch. 5μl (batch) +4 5μl BGB = 50μl.
- .5μm (FL) expired on 01/07/2010.5μl (batch)+45μl BGB = 50μl.
Procedure[2]
- Flow anti-dig 10μl wait for 5 min.
- Flow BGB 50μl wait for 2 min.
- Flow 1.1kb (new)10μl wait for 14 min.
- Flow BGB wait none.
- Flow micro-spheres one in each chamber 10μl wait for 14 min.
- Flow BGB 50μl + seal it with nail polish
Results
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