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(LCM): Preparation and Sectioning of Frozen Tissue Blocks and Purification of RNA from Isolated Cel - http://www.cshprotocols.org/cgi/content/extract/2006/1/pdb.prot4107

Category: Primary Cell Isolation and Culture Protocols: Laser Capture Microdissection Protocols: Tissue Preparation for LCM Protocols

LCM isolates specific cells or tissues from samples mounted on microscope slides. The samples are viewed through a thermoplastic film that is attached to a microcentrifuge tube lid. Localized heat, caused by the application of a laser pulse, fuses the membrane to the cells of interest, which can then be harvested for further analysis. RNA and proteins can be purified from the isolated cells, allowing detailed analysis of gene expression. This protocol is divided into three stages. - [Read (LCM): Preparation and Sectioning of Frozen Tissue Blocks and Purification of RNA from Isolated Cel]

(PI 3-Kinase) Activity Assay - http://www.upstate.com/misc/protocol_detail.q.prot.e.assay.a.name.e.Phosphatidyl_Inositol-3_Kinase_Assay_%28PI3%20Kinase%20Assay%29

Category: Signalling Signal Transduction Protocols

Protocol describing (PI 3-Kinase) activity assay. Includes Preparation of Phosphatidylinositol (PI). - [Read (PI 3-Kinase) Activity Assay]

2-D Polyacrylamide Gel Electrophoresis Protocol - http://cgap-mf.nih.gov/Protocols/DNARNAProteomicAnalysis/Proteomics/2DPage.html

Category: Primary Cell Isolation and Culture Protocols: Laser Capture Microdissection Protocols

Protocol for 2-D polyacrylamide gel electrophoresis. Includes: 50 ml IEF Lysis Buffer; 10X Electrophoresis Running Buffer (10 L); 30% Acrylamide Stock (1 liter); Separating Acrylamide Gel; 50 ml Equilibration Buffer I; 50 ml Equilibration Buffer II; Transfer Buffer; Sample Preparation; Tissue Processing; Reswelling; Prepare Gradient Acrylamide Gel (9-18%); Transfer and Sequencing of Proteins. - [Read 2-D Polyacrylamide Gel Electrophoresis Protocol]

96-well RNA In Situ Hybridization Protocol - http://www.fruitfly.org/about/methods/RNAinsitu.html

Category: PCR Protocols: In-situ PCR Protocols

96-well RNA In Situ Hybridization Protocol. Probe Preparation, Cell Inoculation, PCR, RNA Probe Preparation, etc, Very In-depth Protocol and Method Conditions. Berkeley Drosophila Genome Project. - [Read 96-well RNA In Situ Hybridization Protocol]

A Concise Guide to cArray Hybridization Procedures - DNA Microarray Analysis - http://www.tigr.org/tdb/microarray/conciseguide.html

Category: DNA Microarray Protocols

Hegde et al., Array Fabrication, microarray Printing, Probe Preparation and Hybridization, RNA Extraction, RNA labeling, Data Collection, Normalization, and Analysis.Institute for Genomic Research, Rockville, MD - [Read A Concise Guide to cArray Hybridization Procedures - DNA Microarray Analysis]

A Concise Guide to cDNA Microarray Analysis â€“ II PDF. - http://pga.tigr.org/PDF/BiotechniquesCookbook_II.pdf

Category: DNA Microarray Protocols

Micro Array Fabrication, Probe Preparation and Hybridization, and Data Collection, Normalization and Analysis. Hegde, et al. biotechniques 2000. - [Read A Concise Guide to cDNA Microarray Analysis â€“ II PDF.]

A Sealed Preparation for Long-Term Observations of Cultured Cells - http://www.cshprotocols.org/cgi/content/abstract/2007/1/pdb.prot4660

Category: Microscopy Protocols: Live-Cell Imaging Protocols

This protocol describes a sealed preparation that allows the continuous long-term observation of cultured mammalian cells on upright or inverted microscopes without environmental CO2 control. The preparation allows for optical conditions consistent with high-quality imaging and good cell viability for at least 100 hours. - [Read A Sealed Preparation for Long-Term Observations of Cultured Cells]

A Sealed Preparation for Long-Term Observations of Cultured Cells: Support Slide Construction - http://www.cshprotocols.org/cgi/content/extract/2007/1/pdb.ip26

Category: Microscopy Protocols: Live-Cell Imaging Protocols

Protocol on the details required of support slide construction for a sealed preparation for long-term observations of cultured cells. - [Read A Sealed Preparation for Long-Term Observations of Cultured Cells: Support Slide Construction]

A Simplified System for Rapid Generation of Recombinant Adenoviruses - http://www.coloncancer.org/adeasy/protocol.htm

Category: Viral Culture and Isolation Protocols: Viral Isolation Protocols

A simplified system for rapid generation of recombinant adenoviruses. Includes: Generation of Recombinants in Bacterial Cells; Viral Production in 293 or 911 Cells; Preparation of High Titer Viral Stocks. - [Read A Simplified System for Rapid Generation of Recombinant Adenoviruses]

A Single-step Method for the Simultaneous Preparation of DNA, RNA, and Protein from Cells and Tissue - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot4056

Category: Nucleic Acid Purification Protocols

Protocol for a single-step method for the simultaneous preparation of DNA, RNA, and protein from cells and tissues. The yield of total RNA depends on the tissue or cell source, but it is generally in the range of 4-7 µg/mg starting tissue or 5-10 µg/106 cells. IMPORTANT: Prepare all reagents used in this protocol with Diethyl pyrocarbonate (DEPC)-treated H2O. - [Read A Single-step Method for the Simultaneous Preparation of DNA, RNA, and Protein from Cells and Tissue]

ABI Prism 310 Cycle Sequencing Protocol - http://vosshall.rockefeller.edu/protocols/ABICYCLESEQ.pdf

Category: DNA Protocols: DNA Sequencing Protocols

Protocol for ABI prism 310 cycle sequencing. Includes: Sequencing Reaction; Column Purification; Sample Loading Preparation. - [Read ABI Prism 310 Cycle Sequencing Protocol]

AFLP protocol - http://www.msu.edu/user/hazensam/aflp/AFLPprotocolMSU.html

Category: PCR Protocols: AFLP PCR

Restriction Digestion of Genomic DNA , Adapter preparation, Ligation of Adapters. Pre amplification Reactions. Gel electrophoresis, Silver Staining. AFLP Protocol and procedure. - [Read AFLP protocol]

Agar Plates for Selection of Clones in Bacteria - http://www.le.ac.uk/biology/phh4/bactplte.htm

Category: Microbiology: Bacterial E.Coli Culture Media & Plates

LB Agar: Luria Broth, X-Gal preparation, IPTG and ampicillin. Petri dish preparation. Pat Heslop-Harrison and Trude Schwarzacher, Univ. Leicester - [Read Agar Plates for Selection of Clones in Bacteria]

Agarose Gel Electrophoresis PDF - http://www.uni-tuebingen.de/evoeco/files/methods/Electrophoresis.pdf

Category: Molecular Biology Protocols: Agarose Gel Preparation

Agarose Gel Electrophoresis. Pouring of Agarose Gel, Preparation of samples, Gel electrophoresis, Ethidium bromide staining. - [Read Agarose Gel Electrophoresis PDF]

Agarose Gel Preparation Pictures - http://www.biotech.iastate.edu/publications/ppt_presentations/html/Fingerprinting/AgaroseGelPrep/02.html

Category: General Research Protocols and Methods

Agarose Gel Preparation Pictures. Mike Zeller - [Read Agarose Gel Preparation Pictures]

Agarose Gel Preparation Protocol - http://fp.bio.utk.edu/mycology/Techniques/mt-agarose_gels.htm

Category: Molecular Biology Protocols: Agarose Gel Preparation

Solutions for Agarose gel prepartion, Selection of Appropriate Gel Size and Composition. Mycology Lab University of Tennessee - [Read Agarose Gel Preparation Protocol]

Aggregation of ES Cells with Mouse Morula- Stage Embryos Protocol - http://fds.oup.com/www.oup.co.uk/pdf/pas/13-10-6.pdf

Category: Stem Cell Protocols: Stem Cell Culture Protocols

Protocol for the aggregation of ES cells with mouse morula-stage embryos. Includes: Preparation of aggregation walls; ES cell preparation; Embryo preparation and aggregation. - [Read Aggregation of ES Cells with Mouse Morula- Stage Embryos Protocol]

Aggregation: Production of Chimerae - http://transgenics.ufscc.ufl.edu/TCF05agg_instruct.html

Category: Cell Biology and Cell Culture: ES Stem Cell Biology and Culture: ES Cell Aggregations - Production of Chimerae

Aggregation: Production of Chimerae. Detailed ES cell growth and preparation information for aggregations. Edward W. Scott Univ. Florida Shands Cancer Center - [Read Aggregation: Production of Chimerae]

Amplification of Baculovirus Plaques and Preparation of High Titer Virus Stocks Protocol - http://www.bio.com/protocolstools/protocol.jhtml?id=p608

Category: Cloning Protocols: Baculovirus Protein Expression Protocols

Protocol for amplification of Baculovirus plaques and preparation of high titer virus stocks. Includes: Primary Amplification; Secondary Amplification. - [Read Amplification of Baculovirus Plaques and Preparation of High Titer Virus Stocks Protocol]

An Alternatively Spliced Isoform of t(8;21) Transcript Promotes Leukemogenesis - http://www.natureprotocols.com/2006/08/03/an_alternatively_spliced_isofo.php

Category: Genetics and Genomics: Cancer Genetics Protocols

Describe the methods to identify and quantitate the specific A/E9a transcript in t(8;21) patients samples relative to the AML1-ETO transcript encoding the well known full-length 752 amino acid AML1-ETO protein (AE). Includes: RNA preparation and RT-PCR; Relative quantitation of the AE9a and the AE transcripts. - [Read An Alternatively Spliced Isoform of t(8;21) Transcript Promotes Leukemogenesis]

Annealing, Cloning, and Sequencing of shRNA Linkers into pEN_H1 Plasmids Protocol - http://www.signaling-gateway.org/data/cgi-bin/ProtocolFile.cgi/afcs_PP00000230.pdf?pid=PP00000230

Category: RNAi Technology Protocols

Protocol describes the preparation of an shRNA-expressing Gateway entry vector. This process is preceded by the design and synthesis of target gene-specific sense and antisense oligonucleotides which, when annealed, will constitute an shRNA cassette. - [Read Annealing, Cloning, and Sequencing of shRNA Linkers into pEN_H1 Plasmids Protocol]

Antibiotic solutions - http://molbiol.ru/eng/protocol/01_05.html

Category: Microbiology: Antibiotics Concentration in Media LB

Working concentrations and stock solutions for antibiotics, preparation of stock solutions for Ampicillin, Kanamycin and others. Practical Molecular Biology - [Read Antibiotic solutions]

Antibiotics - http://www.unizh.ch/botinst/Cyto_Website/schneitzLab/Materials/antibiotics.html

Category: Microbiology: Antibiotics Concentration in Media LB

Antibiotic stock solutions preparation, Storage conditions for antibiotics, Working Concentrations for high copy plasmids. Kay Schneitz Univ Zurich. - [Read Antibiotics]

Antibiotics. and Antibiotics Stock Calculator Tools - http://molbiol.edu.ru/eng/protocol/01_05.html

Category: Microbiology: Antibiotics Concentration in Media LB

Working concentrations and stock solutions and which antibiotics are light sensitive. Preparation of stock solutions and calculator tool to calculate concentrations for stock solutions. Practical Molecular Biology. - [Read Antibiotics. and Antibiotics Stock Calculator Tools]

Antibiotics. Preparation and Storage. - http://www.hos.ufl.edu/mooreweb/TissueCulture/class2/Antibiotics%20Sigma.htm

Category: Buffers Media, and Solutions: Bacterial Solutions

Preparation and Storage. Sigma - [Read Antibiotics. Preparation and Storage.]

Articles (11-13 of 13)

Electrotransformation of BMH 81-17mut S for Isolating Site-Directed dsDNA Mutants

This protocol describes the electroporation of the BMH 81-17 mut S strain that is recommended for tranformation of the site directed mutagenesis of dsDNA (See Protocol on Site-Directed Mutagenesis on Double Stranded DNA). BMH 81-17 mut S are a mismatch repair defective (mut S) Escherichia coli strain. The probability that the two mutations will cosegregate during the first round of DNA replication is increased in this strain.

Vector Design Protocol for Transgenic Mice Generation

The protocol gives general considerations for the design of targeting vectors for transgenic mice. The protocol shares tips in the design of knock-out and knock-in vectors and some of their strategies for producing homologously recombined embryonic stem cells.

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