User:Pranav Rathi/Notebook/OT/2013/01/09/DNA-Overstretching Experiments

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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.
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}}
{{#widget:YouTube|id=vKV4CZYMXSw}}{{#widget:YouTube|id=ihl45JssNTA}}
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== Experiment 1 (01/04/2013)==
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Prepare some new buffers and attempt DNA-tethering in H2O and D2O with 265nm beads (bead radius).
 +
 +
===Buffer preparation===
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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''[http://openwetware.org/wiki/User:Pranav_Rathi/Notebook/OT/2012/10/01/Buffer_preparation_for_DNA_overstretching_%26_unzipping_experiments]
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====BGB====
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*H2O
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50mg of BGB + 10ml of H2O 1X POP=5mg/ml BGB 1XPOP H2O 10ml
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*D2O
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50mg of BGB + 10ml of D2O 1X POP=5mg/ml BGB 1XPOP D2O 10ml
 +
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====Antidig====
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same for both H2O and D2O since PBS is in H2O only.
 +
 +
20ul of aliquots + 180ul of PBS H2O=200ul of antidig H2O
 +
 +
===Sample preparation===
 +
 +
 +
 +

Revision as of 18:27, 14 January 2013

Contents

Introduction

This Page contains all the information regarding DNA-Overstretching experiments and results.

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.


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

Sample preparation

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