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Endophytic Microbial Community DNA Extraction from the Plant Phyllosphere

Featured protocol,  Authors: Carlos A. Ruiz-Pérez
Carlos A. Ruiz-PérezAffiliation 1: School of Biological Sciences, Georgia Institute of Technology, Ford Environmental Sciences & Technology Building, Atlanta, USA
Affiliation 2: Molecular Genetics, Corporación Corpogen, Bogotá, Colombia
For correspondence: cruizperez3@gatech.edu
Bio-protocol author page: a4126
 and María Mercedes Zambrano
María Mercedes ZambranoAffiliation: Molecular Genetics, Corporación Corpogen, Bogotá, Colombia
Bio-protocol author page: a4127
date: 2/20/2017, 30 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.2142.

Brief version appeared in Appl Environ Microbiol, Jan 2016
The plant phyllosphere, which represents all plant parts that are above the ground, is considered one of the most extensive ecosystems to be colonized by microorganisms, both at the surface as epiphytes or as endophytes within the plant. These plant-associated microbial communities are reservoirs of microbial diversity and they can be important for plant health. The characterization of microbial communities in diverse plants, such as Espeletia plants that are endemic to the Paramo ecosystem in the Andes Mountains, can shed light regarding possible interactions among microorganisms and microbial functional properties. Obtaining DNA from plant endophytic microbial communities involves various steps to ensure that samples are free of contamination from microorganisms present on the plant surface (epiphytes). Plant leaves are first surface sterilized, cut into pieces, homogenized using glass beads, and then used for DNA extraction using a commercially available kit. DNA samples are then quantified and analyzed using Qubit® 2.0 for use in PCR amplification of 16S rRNA genes.

Production of Guide RNAs in vitro and in vivo for CRISPR Using Ribozymes and RNA Polymerase II Promoters

Featured protocol,  Authors: Tao Zhang*
Tao ZhangAffiliation: National Key Laboratory of Crop Genetic Improvement and National Center of Plant Gene Research (Wuhan), Huazhong Agricultural University, Wuhan, China
Bio-protocol author page: a4181
Yangbin Gao*
Yangbin GaoAffiliation: Section of Cell and Developmental Biology, University of California, San Diego, USA
Bio-protocol author page: a4182
Rongchen Wang
Rongchen WangAffiliation: National Key Laboratory of Crop Genetic Improvement and National Center of Plant Gene Research (Wuhan), Huazhong Agricultural University, Wuhan, China
Bio-protocol author page: a4183
 and Yunde Zhao
Yunde ZhaoAffiliation 1: National Key Laboratory of Crop Genetic Improvement and National Center of Plant Gene Research (Wuhan), Huazhong Agricultural University, Wuhan, China
Affiliation 2: Section of Cell and Developmental Biology, University of California, San Diego, USA
For correspondence: yundezhao@ucsd.edu
Bio-protocol author page: a4184
 (*contributed equally to this work) date: 2/20/2017, 31 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.2148.

Brief version appeared in J Integr Plant Biol, Apr 2014
CRISPR/Cas9-mediated genome editing relies on a guide RNA (gRNA) molecule to generate sequence-specific DNA cleavage, which is a prerequisite for gene editing. Here we establish a method that enables production of gRNAs from any promoters, in any organisms, and in vitro (Gao and Zhao, 2014). This method also makes it feasible to conduct tissue/cell specific gene editing.

Polysome Fractionation to Analyze mRNA Distribution Profiles

Featured protocol,  Authors: Amaresh C. Panda
Amaresh C. PandaAffiliation: Laboratory of Genetics and Genomics, National Institute on Aging, National Institutes of Health, Biomedical Research Center, Baltimore, USA
For correspondence: amarchpanda@gmail.com
Bio-protocol author page: a3875
Jennifer L. Martindale
Jennifer L. Martindale Affiliation: Laboratory of Genetics and Genomics, National Institute on Aging, National Institutes of Health, Biomedical Research Center, Baltimore, USA
Bio-protocol author page: a3880
 and Myriam Gorospe
Myriam GorospeAffiliation: Laboratory of Genetics and Genomics, National Institute on Aging, National Institutes of Health, Biomedical Research Center, Baltimore, USA
Bio-protocol author page: a3881
date: 2/5/2017, 131 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.2126.

Brief version appeared in Nucleic Acids Res, Mar 2016
Eukaryotic cells adapt to changes in external or internal signals by precisely modulating the expression of specific gene products. The expression of protein-coding genes is controlled at the transcriptional and post-transcriptional levels. Among the latter steps, the regulation of translation is particularly important in cellular processes that require rapid changes in protein expression patterns. The translational efficiency of mRNAs is altered by RNA-binding proteins (RBPs) and noncoding (nc)RNAs such as microRNAs (Panda et al., 2014a and 2014b; Abdelmohsen et al., 2014). The impact of factors that regulate selective mRNA translation is a critical question in RNA biology. Polyribosome (polysome) fractionation analysis is a powerful method to assess the association of ribosomes with a given mRNA. It provides valuable information about the translational status of that mRNA, depending on the number of ribosomes with which they are associated, and identifies mRNAs that are not translated (Panda et al., 2016). mRNAs associated with many ribosomes form large polysomes that are predicted to be actively translated, while mRNAs associated with few or no ribosomes are expected to be translated poorly if at all. In sum, polysome fractionation analysis allows the direct determination of translation efficiencies at the level of the whole transcriptome as well as individual mRNAs.

Synthetic Lethality Screens Using RNAi in Combination with CRISPR-based Knockout in Drosophila Cells

Featured protocol,  Authors: Benjamin E. Housden*
Benjamin E. HousdenAffiliation: Harvard Medical School, Boston, USA
For correspondence: bhousden@genetics.med.harvard.edu
Bio-protocol author page: a4035
Hilary E. Nicholson*
Hilary E. NicholsonAffiliation 1: Harvard Medical School, Boston, USA
Affiliation 2: Dana-Farber Cancer Institute, Boston, USA
Bio-protocol author page: a4036
 and Norbert Perrimon
Norbert PerrimonAffiliation 1: Harvard Medical School, Boston, USA
Affiliation 2: Howard Hughes Medical Institute, Boston, USA
For correspondence: perrimon@receptor.med.harvard.edu
Bio-protocol author page: a4037
 (*contributed equally to this work) date: 2/5/2017, 159 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.2119.

Brief version appeared in Sci Signal, Sep 2015
A synthetic lethal interaction is a type of genetic interaction where the disruption of either of two genes individually has little effect but their combined disruption is lethal. Knowledge of synthetic lethal interactions can allow for elucidation of network structure and identification of candidate drug targets for human diseases such as cancer. In Drosophila, combinatorial gene disruption has been achieved previously by combining multiple RNAi reagents. Here we describe a protocol for high-throughput combinatorial gene disruption by combining CRISPR and RNAi. This approach previously resulted in the identification of highly reproducible and conserved synthetic lethal interactions (Housden et al., 2015).

Endophytic Microbial Community DNA Extraction from the Plant Phyllosphere

Authors: Carlos A. Ruiz-Pérez
Carlos A. Ruiz-PérezAffiliation 1: School of Biological Sciences, Georgia Institute of Technology, Ford Environmental Sciences & Technology Building, Atlanta, USA
Affiliation 2: Molecular Genetics, Corporación Corpogen, Bogotá, Colombia
For correspondence: cruizperez3@gatech.edu
Bio-protocol author page: a4126
 and María Mercedes Zambrano
María Mercedes ZambranoAffiliation: Molecular Genetics, Corporación Corpogen, Bogotá, Colombia
Bio-protocol author page: a4127
date: 2/20/2017, 30 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.2142.

[Abstract] The plant phyllosphere, which represents all plant parts that are above the ground, is considered one of the most extensive ecosystems to be colonized by microorganisms, both at the surface as epiphytes or as endophytes within the plant. These plant-associated microbial communities are reservoirs of microbial diversity and they can be important for ...

Production of Guide RNAs in vitro and in vivo for CRISPR Using Ribozymes and RNA Polymerase II Promoters

Authors: Tao Zhang*
Tao ZhangAffiliation: National Key Laboratory of Crop Genetic Improvement and National Center of Plant Gene Research (Wuhan), Huazhong Agricultural University, Wuhan, China
Bio-protocol author page: a4181
Yangbin Gao*
Yangbin GaoAffiliation: Section of Cell and Developmental Biology, University of California, San Diego, USA
Bio-protocol author page: a4182
Rongchen Wang
Rongchen WangAffiliation: National Key Laboratory of Crop Genetic Improvement and National Center of Plant Gene Research (Wuhan), Huazhong Agricultural University, Wuhan, China
Bio-protocol author page: a4183
 and Yunde Zhao
Yunde ZhaoAffiliation 1: National Key Laboratory of Crop Genetic Improvement and National Center of Plant Gene Research (Wuhan), Huazhong Agricultural University, Wuhan, China
Affiliation 2: Section of Cell and Developmental Biology, University of California, San Diego, USA
For correspondence: yundezhao@ucsd.edu
Bio-protocol author page: a4184
 (*contributed equally to this work) date: 2/20/2017, 31 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.2148.

[Abstract] CRISPR/Cas9-mediated genome editing relies on a guide RNA (gRNA) molecule to generate sequence-specific DNA cleavage, which is a prerequisite for gene editing. Here we establish a method that enables production of gRNAs from any promoters, in any organisms, and in vitro (Gao and Zhao, 2014). This method also makes it feasible to conduct tissue/cell ...

Polysome Fractionation to Analyze mRNA Distribution Profiles

Authors: Amaresh C. Panda
Amaresh C. PandaAffiliation: Laboratory of Genetics and Genomics, National Institute on Aging, National Institutes of Health, Biomedical Research Center, Baltimore, USA
For correspondence: amarchpanda@gmail.com
Bio-protocol author page: a3875
Jennifer L. Martindale
Jennifer L. Martindale Affiliation: Laboratory of Genetics and Genomics, National Institute on Aging, National Institutes of Health, Biomedical Research Center, Baltimore, USA
Bio-protocol author page: a3880
 and Myriam Gorospe
Myriam GorospeAffiliation: Laboratory of Genetics and Genomics, National Institute on Aging, National Institutes of Health, Biomedical Research Center, Baltimore, USA
Bio-protocol author page: a3881
date: 2/5/2017, 131 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.2126.

[Abstract] Eukaryotic cells adapt to changes in external or internal signals by precisely modulating the expression of specific gene products. The expression of protein-coding genes is controlled at the transcriptional and post-transcriptional levels. Among the latter steps, the regulation of translation is particularly important in cellular processes that require ...

Synthetic Lethality Screens Using RNAi in Combination with CRISPR-based Knockout in Drosophila Cells

Authors: Benjamin E. Housden*
Benjamin E. HousdenAffiliation: Harvard Medical School, Boston, USA
For correspondence: bhousden@genetics.med.harvard.edu
Bio-protocol author page: a4035
Hilary E. Nicholson*
Hilary E. NicholsonAffiliation 1: Harvard Medical School, Boston, USA
Affiliation 2: Dana-Farber Cancer Institute, Boston, USA
Bio-protocol author page: a4036
 and Norbert Perrimon
Norbert PerrimonAffiliation 1: Harvard Medical School, Boston, USA
Affiliation 2: Howard Hughes Medical Institute, Boston, USA
For correspondence: perrimon@receptor.med.harvard.edu
Bio-protocol author page: a4037
 (*contributed equally to this work) date: 2/5/2017, 159 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.2119.

[Abstract] A synthetic lethal interaction is a type of genetic interaction where the disruption of either of two genes individually has little effect but their combined disruption is lethal. Knowledge of synthetic lethal interactions can allow for elucidation of network structure and identification of candidate drug targets for human diseases such as cancer. ...

Virus Binding and Internalization Assay for Adeno-associated Virus

Authors: Garrett E. Berry
Garrett E. BerryAffiliation 1: Gene Therapy Center, The University of North Carolina at Chapel Hill, Chapel Hill, NC, USA
Affiliation 2: Curriculum in Genetics and Molecular Biology, The University of North Carolina at Chapel Hill, Chapel Hill, NC, USA
For correspondence: berrygar@email.unc.edu
Bio-protocol author page: a3988
 and Longping V. Tse
Longping V. TseAffiliation: Gene Therapy Center, The University of North Carolina at Chapel Hill, Chapel Hill, NC, USA
Bio-protocol author page: a940
date: 1/20/2017, 176 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.2110.

[Abstract] The binding and internalization of adeno-associated virus (AAV) is an important determinant of viral infectivity and tropism. The ability to dissect these two tightly connected cellular processes would allow better understanding and provide insight on virus entry and trafficking. In the following protocol, we describe a quantitative PCR (qPCR) based ...

Dot Blot Analysis of N6-methyladenosine RNA Modification Levels

Authors: Lisha Shen
Lisha ShenAffiliation: Temasek Life Sciences Laboratory and Department of Biological Sciences, National University of Singapore, Singapore
Bio-protocol author page: a4000
Zhe Liang
Zhe Liang Affiliation: Temasek Life Sciences Laboratory and Department of Biological Sciences, National University of Singapore, Singapore
Bio-protocol author page: a4001
 and Hao Yu
Hao YuAffiliation: Temasek Life Sciences Laboratory and Department of Biological Sciences, National University of Singapore, Singapore
For correspondence: dbsyuhao@nus.edu.sg
Bio-protocol author page: a4002
date: 1/5/2017, 247 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.2095.

[Abstract] N6-methyladenosine (m6A) is the most prevalent internal modification of eukaryotic messenger RNA (mRNA). The total amount of m6A can be detected by several methods, such as dot blot analysis using specific m6A antibodies and quantitative liquid chromatography-tandem mass spectrometry (LC-MS/MS) (Fu et al., 2014; Shen et al., 2016). Here we describe ...

Measuring Procaspase-8 and -10 Processing upon Apoptosis Induction

Authors: Sabine Pietkiewicz
Sabine PietkiewiczAffiliation: Translational Inflammation Research, Institute of Experimental Internal Medicine, Otto von Guericke University Magdeburg, Medical Faculty, Magdeburg, Germany
Present address: Medical advisory service of social health insurance, Essen, Germany
Bio-protocol author page: a3934
Clara Wolfe
Clara WolfeAffiliation: Translational Inflammation Research, Institute of Experimental Internal Medicine, Otto von Guericke University Magdeburg, Medical Faculty, Magdeburg, Germany
Bio-protocol author page: a3935
Jörn H. Buchbinder
Jörn H. BuchbinderAffiliation: Translational Inflammation Research, Institute of Experimental Internal Medicine, Otto von Guericke University Magdeburg, Medical Faculty, Magdeburg, Germany
Bio-protocol author page: a3936
 and Inna N. Lavrik
Inna N. LavrikAffiliation 1: Translational Inflammation Research, Institute of Experimental Internal Medicine, Otto von Guericke University Magdeburg, Medical Faculty, Magdeburg, Germany
Affiliation 2: Federal research center Institute of Cytology and Genetics, Novosibirsk, Russia
For correspondence: inna.lavrik@med.ovgu.de
Bio-protocol author page: a3937
date: 1/5/2017, 284 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.2081.

[Abstract] Apoptosis or programmed cell death is important for multicellular organisms to keep cell homeostasis and for the clearance of mutated or infected cells. Apoptosis can be induced by intrinsic or extrinsic stimuli. The first event in extrinsic apoptosis is the formation of the Death-Inducing Signalling Complex (DISC), where the initiator caspases-8 and ...

Fluorescence in situ Localization of Gene Expression Using a lacZ Reporter in the Heterocyst-forming Cyanobacterium Anabaena variabilis

Authors: Brenda S. Pratte
Brenda S. PratteAffiliation: Department of Biology, University of Missouri – St. Louis, St. Louis, MO, USA
Bio-protocol author page: a3944
 and Teresa Thiel
Teresa ThielAffiliation: Department of Biology, University of Missouri – St. Louis, St. Louis, MO, USA
For correspondence: thiel@umsl.edu
Bio-protocol author page: a3945
date: 1/5/2017, 201 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.2084.

[Abstract] One of the most successful fluorescent proteins, used as a reporter of gene expression in many bacterial, plant and animals, is green fluorescent protein and its modified forms, which also function well in cyanobacteria. However, these fluorescent proteins do not allow rapid and economical quantitation of the reporter gene product, as does the popular ...

Efficient AAV-mediated Gene Targeting Using 2A-based Promoter-trap System

Authors: Sivasundaram Karnan
Sivasundaram KarnanAffiliation: Department of Biochemistry, Aichi Medical University School of Medicine, Nagakute, Aichi, Japan
Bio-protocol author page: a3860
Akinobu Ota
Akinobu OtaAffiliation: Department of Biochemistry, Aichi Medical University School of Medicine, Nagakute, Aichi, Japan
Bio-protocol author page: a3861
Yuko Konishi
Yuko KonishiAffiliation: Department of Biochemistry, Aichi Medical University School of Medicine, Nagakute, Aichi, Japan
Bio-protocol author page: a3862
Md Wahiduzzaman
Md WahiduzzamanAffiliation: Department of Biochemistry, Aichi Medical University School of Medicine, Nagakute, Aichi, Japan
Bio-protocol author page: a3863
Shinobu Tsuzuki
Shinobu TsuzukiAffiliation: Department of Biochemistry, Aichi Medical University School of Medicine, Nagakute, Aichi, Japan
Bio-protocol author page: a3864
Yoshitaka Hosokawa
Yoshitaka HosokawaAffiliation: Department of Biochemistry, Aichi Medical University School of Medicine, Nagakute, Aichi, Japan
Bio-protocol author page: a3865
 and Hiroyuki Konishi
Hiroyuki KonishiAffiliation: Department of Biochemistry, Aichi Medical University School of Medicine, Nagakute, Aichi, Japan
For correspondence: hkonishi@aichi-med-u.ac.jp
Bio-protocol author page: a3866
date: 12/20/2016, 333 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.2058.

[Abstract] Adeno-associated virus (AAV)-based targeting vectors have 1-4-log higher gene targeting efficiencies compared with plasmid-based targeting vectors. The efficiency of AAV-mediated gene targeting is further increased by introducing a promoter-trap system into targeting vectors. In addition, we found that the use of ribosome-skipping 2A peptide rather ...

Affinity Pulldown of Biotinylated RNA for Detection of Protein-RNA Complexes

Authors: Amaresh C Panda
Amaresh C PandaAffiliation: Laboratory of Genetics and Genomics, National Institute on Aging, National Institutes of Health, 251 Bayview Blvd., Baltimore, United States
For correspondence: amaresh.panda@nih.gov
Bio-protocol author page: a3875
Jennifer L. Martindale
Jennifer L. Martindale Affiliation: Laboratory of Genetics and Genomics, National Institute on Aging, National Institutes of Health, 251 Bayview Blvd., Baltimore, United States
Bio-protocol author page: a3880
 and Myriam Gorospe
Myriam GorospeAffiliation: Laboratory of Genetics and Genomics, National Institute on Aging, National Institutes of Health, 251 Bayview Blvd., Baltimore, United States
Bio-protocol author page: a3881
date: 12/20/2016, 438 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.2062.

[Abstract] RNA-binding proteins (RBPs) have recently emerged as crucial players in the regulation of gene expression. The interactions of RBPs with target mRNAs control the levels of gene products by altering different regulatory steps, including pre-mRNA splicing and maturation, nuclear mRNA export, and mRNA stability and translation (Glisovic et al., 2008). ...
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[Bio101] Plasmid DNA Extraction from E. coli Using Alkaline Lysis Method

Author: Fanglian He
Fanglian HeAffiliation: Department of Biology, University of Pennsylvania, Philadelphia, USA
For correspondence: fanglian09@gmail.com
Bio-protocol author page: a9
date: 2/5/2011, 82765 views, 31 Q&A
DOI: https://doi.org/10.21769/BioProtoc.30.

[Abstract] This is a quick and efficient way to extract E. coli plasmid DNA without using commercial kits....

[Bio101] E. coli Genomic DNA Extraction Updates
The author made some updates (highlighted in blue) to the protocol on 09/12/2016.

Author: Fanglian He
Fanglian HeAffiliation: Department of Biology, University of Pennsylvania, Philadelphia, USA
Bio-protocol author page: a9
date: 7/20/2011, 75542 views, 46 Q&A
DOI: https://doi.org/10.21769/BioProtoc.97.

[Abstract] This protocol uses phenol/chloroform method to purify genomic DNA without using commercial kits....

[Bio101] DNA Molecular Weight Calculation

Author: Fanglian He date: 3/20/2011, 33770 views, 7 Q&A
DOI: https://doi.org/10.21769/BioProtoc.46.

[Abstract] This method is to roughly estimate DNA molecular weight. One of its applications is to calculate the ratio of vector to insert in a ligation reaction (please see Standard DNA Cloning protocol).
Anhydrous molecular weight of each nucleotide is (see reference 1):
A= 313.21
T= 304.2
C= 289.18
G=329.21
For rough ...





[Bio101] Infiltration of Nicotiana benthamiana Protocol for Transient Expression via Agrobacterium

Author: Xiyan Li
Xiyan LiAffiliation: Department of Genetics, Stanford University, Stanford, USA
For correspondence: lixiyan@stanford.edu
Bio-protocol author page: a13
date: 7/20/2011, 30176 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.95.

[Abstract] Transient expression in tobacco plant (Nicotiana benthamiana) is used to determine the subcellular location of a protein of interest when tagged with a reporter such as green fluorescent protein (GFP), or to mass produce proteins without making transgenic plants. The root tumor bacteria, Agrobacteria, ...

[Bio101] Calcium Phosphate Transfection of Eukaryotic Cells

Author: Yanling Chen
Yanling ChenAffiliation: Department of Immunology, The Scripps Research Institute, La Jolla, USA
For correspondence: ylchen@scripps.edu
Bio-protocol author page: a27
date: 2/5/2012, 26591 views, 2 Q&A
DOI: https://doi.org/10.21769/BioProtoc.86.

[Abstract] Transfection of DNA into cells is an indispensible protocol in molecular biology. While plenty of lipid-based transfection reagents are commercially available nowadays, a quick, simple, efficient and inexpensive method is to transfect eukaryotic cells via calcium phosphate co-precipitation with DNA ...

C2C12 Myoblasts

Author: Lili Jing
Lili JingAffiliation: Department of Cell and Molecular Biology, University of Pennsylvania, Philadelphia, USA
For correspondence: lilijingcn@gmail.com
Bio-protocol author page: a38
date: 5/20/2012, 26258 views, 11 Q&A
DOI: https://doi.org/10.21769/BioProtoc.172.

[Abstract] C2C12 myoblasts are commonly used in biomedical laboratories as an in vitro system to study muscle development and differentiation. This protocol explains the basic procedures of culture, transfection and differentiation of C2C12 myoblast cells....

[Bio101] Standard DNA Cloning

Author: Fanglian He date: 4/5/2011, 23346 views, 4 Q&A
DOI: https://doi.org/10.21769/BioProtoc.52.

[Abstract] This protocol describes general cloning steps from preparation of both vector and insert DNA to the ligation reaction....

[Bio101] A General EMSA (Gel-shift) Protocol

Author: Ran Chen
Ran ChenAffiliation: Department of Genetics, Stanford University, Stanford, USA
For correspondence: rcchen@jfkbio.com
Bio-protocol author page: a34
date: 2/5/2011, 23332 views, 3 Q&A
DOI: https://doi.org/10.21769/BioProtoc.24.

[Abstract] An electrophoretic mobility shift assay (EMSA), also referred to as mobility shift electrophoresis, a gel shift assay, gel mobility shift assay, band shift assay, or gel retardation assay, is a common technique used to study protein-DNA or protein-RNA interactions. The control lane (the DNA/RNA probe ...

[Bio101] Lentivirus Production

Author: Nabila Aboulaich date: 3/5/2011, 22389 views, 6 Q&A
DOI: https://doi.org/10.21769/BioProtoc.39.

[Abstract] Lentivirus is a common tool used to introduce a gene into mammalian or other animal cells.This protocol is to produce lentivirus stocks from hairpin-pLKO.1 plasmid....

In vitro Protein Kinase Assay

Author: Yuehua Wei
Yuehua WeiAffiliation: Department of Pharmacology, Cancer Institute of New Jersey, UMDNJ Robert Wood Johnson Medical School, Piscataway, USA
For correspondence: weiyh.sjtu.edu@gmail.com
Bio-protocol author page: a49
date: 6/5/2012, 21038 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.212.

[Abstract] This protocol will describe experimental procedures for an in vitro kinase assay of the yeast protein kinase Sch9. This protocol can be tailored to detect kinase activity of other yeast protein kinase....
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