User:Pranav Rathi/Notebook/OT/2012/10/12/Sample preparation for DNA overstretching & unzipping experiments

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(Chamber preparation)
 
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The page discusses the protocol we use to prepare DNA-tethering, overstretching and unzipping samples. A standard protocol is same for all three; a specific but similar protocol is adopted depending on the sample and its constituents. I also discuss protocol for sample used to calibrate optical tweezers; stuck and free bead. A standard sample looks like slide.1 in slide-share: It can be single or double chamber. A sample preparation can be divided into two parts: Chamber (single or double) preparation and Tethering-protocol. Sample preparation-procedure discussed here is standard, specific but similar procedure is used depending on the requirements of the sample.
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In all my DNA and calibration experiments I use this procedure and it works really well for me. It is simple, quick, versatile and very effective.
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=Sample preparation=
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Sample preparation can be divided in chamber preparation and tethering-protocol.
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==Chamber preparation==
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A double chamber slide preparation is discussed here with components:
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* Slide (by VWR micro slides, cat# 48300-025 [http://vwrlabshop.com/vwr-174-plain-and-frosted-micro-slides/p/0025555/search/48300-025/])
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* Cover glass; 18X18mm,.15mm thick (by VWR micro cover glass, cat# 48366-045[http://vwrlabshop.com/vwr-micro-cover-glasses-square-no1/p/0010921/search/48366-045/])
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* Double sided tape: This is a double sided tape by scotch; cheep but good enough for quality samples.
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* Tweezers.
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* Roller ( a solid aluminium cylinder to squeezeout the air bubbles between the tape and glass-slide).
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* Double razer
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Current revision

The page discusses the protocol we use to prepare DNA-tethering, overstretching and unzipping samples. A standard protocol is same for all three; a specific but similar protocol is adopted depending on the sample and its constituents. I also discuss protocol for sample used to calibrate optical tweezers; stuck and free bead. A standard sample looks like slide.1 in slide-share: It can be single or double chamber. A sample preparation can be divided into two parts: Chamber (single or double) preparation and Tethering-protocol. Sample preparation-procedure discussed here is standard, specific but similar procedure is used depending on the requirements of the sample.

In all my DNA and calibration experiments I use this procedure and it works really well for me. It is simple, quick, versatile and very effective.


Sample preparation

Sample preparation can be divided in chamber preparation and tethering-protocol.

Chamber preparation

A double chamber slide preparation is discussed here with components:

  • Slide (by VWR micro slides, cat# 48300-025 [1])
  • Cover glass; 18X18mm,.15mm thick (by VWR micro cover glass, cat# 48366-045[2])
  • Double sided tape: This is a double sided tape by scotch; cheep but good enough for quality samples.
  • Tweezers.
  • Roller ( a solid aluminium cylinder to squeezeout the air bubbles between the tape and glass-slide).
  • Double razer








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