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The resolution of conventional magnetic tweezers is limited by the Brown motion of magnetic beads. When the force is lower than ～10 pN, the resolution of magnetic tweezers decreases significantly because of the increased Brown motion. To improve the resolution of magnetic tweezers under low forces, we combine the total internal reflection fluorescence techniques with magnetic tweezers, and design a novel single molecule connection: magnetic bead-DNA linker-fluorescent bead-single molecule. With the improved magnetic tweezers, we study the folding dynamics of a DNA hairpin. The results reveal that a nanometer-scale resolution is obtained. By analyzing these results, we calibrate the penetration depth of the total internal reflection field. Finally, we investigate the unwinding dynamics of a BLM helicase core protein. Some preliminary results of the helicase unwinding experiments confirm the practicability of the improved magnetic tweezers in the field of single molecular research.
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Keywords:
- magnetic tweezers /
- total internal reflection fluorescence /
- DNA hairpin /
- helicase
[1] Smith S B, Finazi L, Bustamante C 1992 Science 258 1122
[2] Cluzel P, Lebrun A, Heller C, Lavery R, Viovy J, Chatenay C, Caron F 1996 Science 271 792
[3] Dessinges M N, Lionnet T, Xi X G, Bensimon D, Croquette V 2004 Proc. Natl. Acad. Sci. USA 101 6439
[4] Johnson D S, Bai L, Smith B Y, Patel S S, Wang M D 2007 Cell 129 1299
[5] Perkings T T, Li H W, Dalal R V, Gelles J, Block S M 2004 Biophys. J. 86 1640
[6] Marsden S, Nardelli M, Linder P, Mccarthy J E 2006 J. Mol. Biol. 361 327
[7] Myong S, Bruno M M, Pyle A M, Ha T 2007 Science 317 513
[8] Theissen B, Karow A R, Kohler J, Gubaev A, Klostermeier D 2008 Proc. Natl. Acad. Sci. USA 105 548
[9] Smith S B, Cui Y, Bustamante C 1996 Science 271 795
[10] Sun B, Wei K J, Zhang B, Zhang X H, Dou S X, Li M, Xi X G 2008 EMBO J. 27 3279
[11] Neuman K C, Nagy A 2008 Nat. Methods 5 491
[12] Marko J F, Siggia E D 1995 Macromolecules 28 8759
[13] Lionnet T, Spiering M M, Benkovic S J, Bensimon D, Croquette V 2007 Natl. Acad. Sci. USA 104 19790
[14] Ran S Y 2012 Acta Phys. Sin. 61 170503 (in Chinese) [冉诗勇 2012 61 170503]
[15] Hohng S, Zhou R, Nahas M K, Yu J, Schulten K, Lilley D M, Ha T 2007 Science 318 279
[16] Lee M, Kim S H, Hong S C 2010 Proc. Natl. Acad. Sci. USA 107 4985
[17] Luzzietti N, Brutzer H, Klaue D, Schwarz F W, Staroske W, Clausing S, Seidel R 2010 Nucl. Acids Res. 39 e15
[18] Guo R B, Rigolet P, Zargarian L, Fermandjian S, Xi X G 2005 Nucl. Acids Res. 33 3109
[19] Yang Y, Dou S X, Xu Y N, Bazeille N, Wang P Y, Rigolet P, Xu H Q, Xi X G 2010 Biochemistry 49 656
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[1] Smith S B, Finazi L, Bustamante C 1992 Science 258 1122
[2] Cluzel P, Lebrun A, Heller C, Lavery R, Viovy J, Chatenay C, Caron F 1996 Science 271 792
[3] Dessinges M N, Lionnet T, Xi X G, Bensimon D, Croquette V 2004 Proc. Natl. Acad. Sci. USA 101 6439
[4] Johnson D S, Bai L, Smith B Y, Patel S S, Wang M D 2007 Cell 129 1299
[5] Perkings T T, Li H W, Dalal R V, Gelles J, Block S M 2004 Biophys. J. 86 1640
[6] Marsden S, Nardelli M, Linder P, Mccarthy J E 2006 J. Mol. Biol. 361 327
[7] Myong S, Bruno M M, Pyle A M, Ha T 2007 Science 317 513
[8] Theissen B, Karow A R, Kohler J, Gubaev A, Klostermeier D 2008 Proc. Natl. Acad. Sci. USA 105 548
[9] Smith S B, Cui Y, Bustamante C 1996 Science 271 795
[10] Sun B, Wei K J, Zhang B, Zhang X H, Dou S X, Li M, Xi X G 2008 EMBO J. 27 3279
[11] Neuman K C, Nagy A 2008 Nat. Methods 5 491
[12] Marko J F, Siggia E D 1995 Macromolecules 28 8759
[13] Lionnet T, Spiering M M, Benkovic S J, Bensimon D, Croquette V 2007 Natl. Acad. Sci. USA 104 19790
[14] Ran S Y 2012 Acta Phys. Sin. 61 170503 (in Chinese) [冉诗勇 2012 61 170503]
[15] Hohng S, Zhou R, Nahas M K, Yu J, Schulten K, Lilley D M, Ha T 2007 Science 318 279
[16] Lee M, Kim S H, Hong S C 2010 Proc. Natl. Acad. Sci. USA 107 4985
[17] Luzzietti N, Brutzer H, Klaue D, Schwarz F W, Staroske W, Clausing S, Seidel R 2010 Nucl. Acids Res. 39 e15
[18] Guo R B, Rigolet P, Zargarian L, Fermandjian S, Xi X G 2005 Nucl. Acids Res. 33 3109
[19] Yang Y, Dou S X, Xu Y N, Bazeille N, Wang P Y, Rigolet P, Xu H Q, Xi X G 2010 Biochemistry 49 656
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