13: Biotechnology Lab Protocols
- Page ID
- 138393
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\(\newcommand{\avec}{\mathbf a}\) \(\newcommand{\bvec}{\mathbf b}\) \(\newcommand{\cvec}{\mathbf c}\) \(\newcommand{\dvec}{\mathbf d}\) \(\newcommand{\dtil}{\widetilde{\mathbf d}}\) \(\newcommand{\evec}{\mathbf e}\) \(\newcommand{\fvec}{\mathbf f}\) \(\newcommand{\nvec}{\mathbf n}\) \(\newcommand{\pvec}{\mathbf p}\) \(\newcommand{\qvec}{\mathbf q}\) \(\newcommand{\svec}{\mathbf s}\) \(\newcommand{\tvec}{\mathbf t}\) \(\newcommand{\uvec}{\mathbf u}\) \(\newcommand{\vvec}{\mathbf v}\) \(\newcommand{\wvec}{\mathbf w}\) \(\newcommand{\xvec}{\mathbf x}\) \(\newcommand{\yvec}{\mathbf y}\) \(\newcommand{\zvec}{\mathbf z}\) \(\newcommand{\rvec}{\mathbf r}\) \(\newcommand{\mvec}{\mathbf m}\) \(\newcommand{\zerovec}{\mathbf 0}\) \(\newcommand{\onevec}{\mathbf 1}\) \(\newcommand{\real}{\mathbb R}\) \(\newcommand{\twovec}[2]{\left[\begin{array}{r}#1 \\ #2 \end{array}\right]}\) \(\newcommand{\ctwovec}[2]{\left[\begin{array}{c}#1 \\ #2 \end{array}\right]}\) \(\newcommand{\threevec}[3]{\left[\begin{array}{r}#1 \\ #2 \\ #3 \end{array}\right]}\) \(\newcommand{\cthreevec}[3]{\left[\begin{array}{c}#1 \\ #2 \\ #3 \end{array}\right]}\) \(\newcommand{\fourvec}[4]{\left[\begin{array}{r}#1 \\ #2 \\ #3 \\ #4 \end{array}\right]}\) \(\newcommand{\cfourvec}[4]{\left[\begin{array}{c}#1 \\ #2 \\ #3 \\ #4 \end{array}\right]}\) \(\newcommand{\fivevec}[5]{\left[\begin{array}{r}#1 \\ #2 \\ #3 \\ #4 \\ #5 \\ \end{array}\right]}\) \(\newcommand{\cfivevec}[5]{\left[\begin{array}{c}#1 \\ #2 \\ #3 \\ #4 \\ #5 \\ \end{array}\right]}\) \(\newcommand{\mattwo}[4]{\left[\begin{array}{rr}#1 \amp #2 \\ #3 \amp #4 \\ \end{array}\right]}\) \(\newcommand{\laspan}[1]{\text{Span}\{#1\}}\) \(\newcommand{\bcal}{\cal B}\) \(\newcommand{\ccal}{\cal C}\) \(\newcommand{\scal}{\cal S}\) \(\newcommand{\wcal}{\cal W}\) \(\newcommand{\ecal}{\cal E}\) \(\newcommand{\coords}[2]{\left\{#1\right\}_{#2}}\) \(\newcommand{\gray}[1]{\color{gray}{#1}}\) \(\newcommand{\lgray}[1]{\color{lightgray}{#1}}\) \(\newcommand{\rank}{\operatorname{rank}}\) \(\newcommand{\row}{\text{Row}}\) \(\newcommand{\col}{\text{Col}}\) \(\renewcommand{\row}{\text{Row}}\) \(\newcommand{\nul}{\text{Nul}}\) \(\newcommand{\var}{\text{Var}}\) \(\newcommand{\corr}{\text{corr}}\) \(\newcommand{\len}[1]{\left|#1\right|}\) \(\newcommand{\bbar}{\overline{\bvec}}\) \(\newcommand{\bhat}{\widehat{\bvec}}\) \(\newcommand{\bperp}{\bvec^\perp}\) \(\newcommand{\xhat}{\widehat{\xvec}}\) \(\newcommand{\vhat}{\widehat{\vvec}}\) \(\newcommand{\uhat}{\widehat{\uvec}}\) \(\newcommand{\what}{\widehat{\wvec}}\) \(\newcommand{\Sighat}{\widehat{\Sigma}}\) \(\newcommand{\lt}{<}\) \(\newcommand{\gt}{>}\) \(\newcommand{\amp}{&}\) \(\definecolor{fillinmathshade}{gray}{0.9}\)- 13.1: Lab Technique - Pipetting
- This page introduces some basic pipetting techniques, such as pipetting with a serological pipette and using a micropipette
- 13.2: Lab Technique - Polymerase Chain Reaction (PCR)
- The page outlines how DNA can be amplified through semi-quantitative Polymerase Chain Reaction (PCR) and quantitated through quantitative PCR (qPCR).
- 13.3: Lab Technique - Agarose Gel Electrophoresis of DNA
- This page outlines how DNA fragments can be separated by size using agarose gel electrophoresis
- 13.4: Lab Technique - Restriction Digest of DNA
- This page outlines how DNA can be digested using restriction enzymes. A protocol for a double digest is outlined.
- 13.5: Lab Technique - Ligation of Insert DNA into a Plasmid
- This page outlines how DNA can be inserted into a bacterial plasmid through a ligation reaction.
- 13.6: Lab Technique - Bacterial Transformation
- This page outlines how DNA can be introduced into bacteria through chemical and electrical transformation.
- 13.7: Lab Technique - Isolation of DNA from Bacteria (The "Mini-Prep")
- This page outlines how recombinant plasmids can be isolated from transformed bacteria.
- 13.8: Lab Technique - Quantitation of DNA
- This page outlines how DNA can be quantitated through UV spectroscopy.
- 13.9: Lab Technique - DNA Sequencing
- This page outlines how DNA can be sequenced through the Sanger method.
- 13.10: Lab Technique - Liposomal transfection of DNA into mammalian cells
- This page outlines how DNA can be introduced into mammalian cells through liposomal transfection.
- 13.11: Lab Technique - SDS-PAGE
- This page outlines how proteins can be separated by size through SDS-Polyacrylamide Gel Electrophoresis (SDS-PAGE).
- 13.12: Lab Technique - Western Blotting
- This page outlines how protein expression can be confirmed through immunoblotting (i.e., Western blotting).
- 13.13: Lab Technique - Cell Culture
- This page outlines several common protocols used in cell culture, including harvesting cells for subculture, counting cells, and cryostoring cells.
- 13.14: Lab Technique - Light Microscopy
- This page details some basic microscopy protocols that are used with a compound light microscope, including oil immersion and indirect immunofluorescence.
- 13.15: Bacterial Identification Methods
- This page outlines some common bacterial staining protocols, including a simple stain with methylene blue and the differential staining protocols, Gram staining, acid-fast staining, and endospore staining. In addition, protocols for preparing bacterial smears and plates are given.
- 13.16: Basics of Bacterial Culture
- This page provides an overview of bacterial culture techniques, including the environmental conditions that influence bacterial growth, protocols for preparing different types of culture media (broth, slants, agar plates). It also outlines proper aseptic techniques for inoculating media and instructions for creating streak plates to isolate individual bacterial colonies.