A general method for manipulating DNA sequences from any organism with optical tweezers | Zendy

Derek N. Fuller | Zendy; Gregory J. Gemmen | Zendy; John Peter Rickgauer | Zendy; Aurélie Dupont | Zendy; Rachel Millin | Zendy; Pierre Recouvreux | Zendy; Douglas E. Smith | Zendy

AI Assistant Blog Pricing

Home ZAIA Blog

Open Access

A general method for manipulating DNA sequences from any organism with optical tweezers

Author(s) -

Derek N. Fuller,

Gregory J. Gemmen,

John Peter Rickgauer,

Aurélie Dupont,

Rachel Millin,

Pierre Recouvreux,

Douglas E. Smith

Publication year - 2006

Publication title -

nucleic acids research

Language(s) - English

Resource type - Journals

SCImago Journal Rank - 9.008

H-Index - 537

eISSN - 1362-4954

pISSN - 0305-1048

DOI - 10.1093/nar/gnj016

Subject(s) - dna , optical tweezers , biology , ionic strength , biophysics , magnetic tweezers , function (biology) , tweezers , biological system , molecule , genomic dna , dna sequencing , genome , computational biology , base pair , genetics , gene , chemistry , physics , optics , organic chemistry , aqueous solution

Mechanical manipulation of single DNA molecules can provide novel information about DNA properties and protein-DNA interactions. Here we describe and characterize a useful method for manipulating desired DNA sequences from any organism with optical tweezers. Molecules are produced from either genomic or cloned DNA by PCR using labeled primers and are tethered between two optically trapped microspheres. We demonstrate that human, insect, plant, bacterial and viral sequences ranging from approximately 10 to 40 kilobasepairs can be manipulated. Force-extension measurements show that these constructs exhibit uniform elastic properties in accord with the expected contour lengths for the targeted sequences. Detailed protocols for preparing and manipulating these molecules are presented, and tethering efficiency is characterized as a function of DNA concentration, ionic strength and pH. Attachment strength is characterized by measuring the unbinding time as a function of applied force. An alternative stronger attachment method using an amino-carboxyl linkage, which allows for reliable DNA overstretching, is also described.

The content you want is available to Zendy users.

Already have an account? Click here to sign in.

Having issues? You can contact us here

Accelerating Research