Laboratory methods in enzymology : cell, lipid and carbohydrate / edited by Jon Lorsch.

Contributor(s): Lorsch, Jon [editor.]
Material type: TextTextSeries: Methods in enzymology: v. 533.Publisher: San Diego, CA : Academic Press, 2013Copyright date: ©2013Description: 1 online resource (lix, 347 pages) : illustrationsContent type: text Media type: computer Carrier type: online resourceOther title: Cell, lipid and carbohydrateSubject(s): Biotechnology -- Methodology | Molecular biology -- Methodology | Carbohydrates -- Laboratory manuals | Cells -- Laboratory manuals | Lipids -- Laboratory manuals | Biotechnology -- Methodology | Carbohydrates | Cells | Lipids | Molecular biology -- Methodology | Biotechnology -- methods | Carbohydrates -- analysis | Molecular Biology -- methodsGenre/Form: Laboratory manuals. | Ebook. | Ebook. | Electronic books. Additional physical formats: Print version:: Laboratory methods in enzymology.DDC classification: 572/.7 LOC classification: QP601 | .L32 2013ebOnline resources: Click here to access online
Contents:
Section I. Cell protocols : basic microbiological techniques. Pouring agar plates and streaking or spreading to isolate individual colonies / Elizabeth Eyler ; Storage of bacteria and yeast / Cheryl M. Koh -- Section II. Cell protocols : cellular fractionation. Cellular fractionation : mammalian cells / Joerg von Hagen and Uwe Michelsen ; Cellular fractionation : yeast cells / Joerg von Hagen and Uwe Michelsen -- Section III. Cell protocols : Knock-outs and gene replacements. Yeast-gene replacement using PCR products / Megan Bergkessel, Christine Guthrie, and John Abelson ; Measuring RNAi knockdown using qPCR / Stuart Milstein, Michael Nguyen, Rachel Meyers, and Antonin de Fougerolles ; Recombineering : using drug cassettes to knock out genes in vivo / James A. Sawitzke, Lynn C. Thomason, Mikhail Bubunenko, Xintian Li, Nina Costantino, and Donald L. Court ; Gene knockouts, in vivo site-directed mutagenesis and other modifications using the delitto perfetto system in Saccharomyces cerevisiae / Samantha Stuckey and Francesca Storici ; Gene targeting and site-specific recombination in mouse ES cells / Konstantinos Anastassiadis, Frank Schnütgen, Harald von Melchner, and A. Francis Stewart ; Recombineering : highly efficient in vivo genetic engineering using single-strand oligos / James A. Sawitzke, Lynn C. Thomason, Mikhail Bubunenko, Xintian Li, Nina Costantino, and Donald L. Court -- Section IV. Cell protocols : microbiological media. Growth media for E. coli / Juliane C. Lessard ; Saccharomyces cerevisiae growth media / Jessica S. Dymond -- Section V. Cell protocols : preparation of slides for microscopy. Immunohistochemistry on freely floating fixed tissue sections / Julia Bachman ; Preparation of slides for microscopy from frozen tissue sections / Julia Bachman ; Preparation of formalin-fixed paraffin-embedded tissue for immunohistochemistry / Kirstie Canene-Adams ; Preparation of cells for microscopy using cytospin / Cheryl M. Koh ; Preparation of cells for microscopy using chamber slides and coverslips / Cheryl M. Koh ; Preparation of cells for microscopy using 'cell blocks' / Cheryl M. Koh -- Section VI. Lipid and carbohydrate protocols. Enzymatic deglycosylation of glycoproteins / Min-Sung Kim and Dan Leahy -- Section VII. Lipid and carbohydrate protocol : making vesicles and micelles. Preparation of fatty acid or phospholipid vesicles by thin-film rehydration / Ting F. Zhu, Itay Budin, and Jack W. Szostak ; Vesicle extrusion through polycarbonate track-etched membranes using a hand-held mini-extruder / Ting F. Zhu, Itay Budin, and Jack W. Szostak ; Preparation of fatty acid micelles / Ting F. Zhu, Itay Budin, and Jack W. Szostak -- Section VIII. Other protocols. Reverse-phase HPLC analysis and purification of small molecules / Kirstie Canene-Adams ; Thin layer chromatography / Marina Santiago and Scott Strobel ; ATP and GTP hydrolysis assays (TLC) / Vaishnavi Rajagopal and Jon R. Lorsch.
Summary: These volumes of Methods in Enzymology contain the protocols that made up the on-line Methods Navigator. Our philosophy when we selected the protocols to include in the Navigator was that they should be for techniques useful in any biomedical laboratory, regardless of the system the lab studies. Each protocol was written by researchers who use the technique routinely, and in many cases by the people who actually developed the procedure in the first place. The protocols are very detailed and contain recipes for the necessary buffers and reagents, as well as flow-charts outlining the steps involved. Many of the chapters have accompanying videos demonstrating key parts of the procedures. The volumes are broken into distinct areas: DNA methods; Cell-based methods; lipid, carbohydrate and miscellaneous methods; RNA methods; protein methods. Our goal is that these protocols will be useful for everyone in the lab, from undergraduates and rotation students to seasoned post-doctoral fellows. We hope that these volumes will become dog-eared and well-worn in your laboratory, either physically or electronically.
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Includes bibliographical references and index.

Print version record.

Section I. Cell protocols : basic microbiological techniques. Pouring agar plates and streaking or spreading to isolate individual colonies / Elizabeth Eyler ; Storage of bacteria and yeast / Cheryl M. Koh -- Section II. Cell protocols : cellular fractionation. Cellular fractionation : mammalian cells / Joerg von Hagen and Uwe Michelsen ; Cellular fractionation : yeast cells / Joerg von Hagen and Uwe Michelsen -- Section III. Cell protocols : Knock-outs and gene replacements. Yeast-gene replacement using PCR products / Megan Bergkessel, Christine Guthrie, and John Abelson ; Measuring RNAi knockdown using qPCR / Stuart Milstein, Michael Nguyen, Rachel Meyers, and Antonin de Fougerolles ; Recombineering : using drug cassettes to knock out genes in vivo / James A. Sawitzke, Lynn C. Thomason, Mikhail Bubunenko, Xintian Li, Nina Costantino, and Donald L. Court ; Gene knockouts, in vivo site-directed mutagenesis and other modifications using the delitto perfetto system in Saccharomyces cerevisiae / Samantha Stuckey and Francesca Storici ; Gene targeting and site-specific recombination in mouse ES cells / Konstantinos Anastassiadis, Frank Schnütgen, Harald von Melchner, and A. Francis Stewart ; Recombineering : highly efficient in vivo genetic engineering using single-strand oligos / James A. Sawitzke, Lynn C. Thomason, Mikhail Bubunenko, Xintian Li, Nina Costantino, and Donald L. Court -- Section IV. Cell protocols : microbiological media. Growth media for E. coli / Juliane C. Lessard ; Saccharomyces cerevisiae growth media / Jessica S. Dymond -- Section V. Cell protocols : preparation of slides for microscopy. Immunohistochemistry on freely floating fixed tissue sections / Julia Bachman ; Preparation of slides for microscopy from frozen tissue sections / Julia Bachman ; Preparation of formalin-fixed paraffin-embedded tissue for immunohistochemistry / Kirstie Canene-Adams ; Preparation of cells for microscopy using cytospin / Cheryl M. Koh ; Preparation of cells for microscopy using chamber slides and coverslips / Cheryl M. Koh ; Preparation of cells for microscopy using 'cell blocks' / Cheryl M. Koh -- Section VI. Lipid and carbohydrate protocols. Enzymatic deglycosylation of glycoproteins / Min-Sung Kim and Dan Leahy -- Section VII. Lipid and carbohydrate protocol : making vesicles and micelles. Preparation of fatty acid or phospholipid vesicles by thin-film rehydration / Ting F. Zhu, Itay Budin, and Jack W. Szostak ; Vesicle extrusion through polycarbonate track-etched membranes using a hand-held mini-extruder / Ting F. Zhu, Itay Budin, and Jack W. Szostak ; Preparation of fatty acid micelles / Ting F. Zhu, Itay Budin, and Jack W. Szostak -- Section VIII. Other protocols. Reverse-phase HPLC analysis and purification of small molecules / Kirstie Canene-Adams ; Thin layer chromatography / Marina Santiago and Scott Strobel ; ATP and GTP hydrolysis assays (TLC) / Vaishnavi Rajagopal and Jon R. Lorsch.

These volumes of Methods in Enzymology contain the protocols that made up the on-line Methods Navigator. Our philosophy when we selected the protocols to include in the Navigator was that they should be for techniques useful in any biomedical laboratory, regardless of the system the lab studies. Each protocol was written by researchers who use the technique routinely, and in many cases by the people who actually developed the procedure in the first place. The protocols are very detailed and contain recipes for the necessary buffers and reagents, as well as flow-charts outlining the steps involved. Many of the chapters have accompanying videos demonstrating key parts of the procedures. The volumes are broken into distinct areas: DNA methods; Cell-based methods; lipid, carbohydrate and miscellaneous methods; RNA methods; protein methods. Our goal is that these protocols will be useful for everyone in the lab, from undergraduates and rotation students to seasoned post-doctoral fellows. We hope that these volumes will become dog-eared and well-worn in your laboratory, either physically or electronically.

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