User:Pranav Rathi/Notebook/OT/2013/01/09/DNA-Overstretching Experiments: Difference between revisions
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=Experiments= | =Experiments= |
Revision as of 15:29, 14 January 2013
Introduction
This Page contains all the information regarding DNA-Overstretching experiments and results.
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Proof of DNA-tethering
Two video presents good DNA-tethers in H2O and D2O. For some reason D2O tethers look little better. Each moving bead has a ds-DNA tether which is roughly 1500nm long (4.4kb; 110ng/ml; 02/11/11). The bead size is 530nm diameter. This sample is prepared in water and heavy water for DNA-overstretching experiments which i will discuss below. {{#widget:YouTube|id=vKV4CZYMXSw}}{{#widget:YouTube|id=ihl45JssNTA}}
Experiments
Experiment 1 (01/04/2013)
Prepare some new buffers and attempt DNA-tethering in H2O and D2O with 265nm beads (bead radius).
Buffer preparation
BGB is good only for few weeks and it has been over 2 months since i made it last, so i am going to make new BGB and antidig. For more information go to Buffer preparation for DNA overstretching and unzippingexperiments[1]
BGB
- H2O
50mg of BGB + 10ml of H2O 1X POP=5mg/ml BGB 1XPOP H2O 10ml
- D2O
50mg of BGB + 10ml of D2O 1X POP=5mg/ml BGB 1XPOP D2O 10ml
Antidig
same for both H2O and D2O since PBS is in H2O only.
20ul of aliquots + 180ul of PBS H2O=200ul of antidig H2O