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A Novel Method for Simultaneous Anterograde and Retrograde Labeling of Spinal Cord Motor Tracts - http://www.jhc.org/cgi/content/full/49/9/1111

Category: Neuroscience Protocols: Neuronal Stains Protocols

Protocol for a method for simultaneous anterograde and retrograde labeling of spinal cord motor tracts in the same animal. - [Read A Novel Method for Simultaneous Anterograde and Retrograde Labeling of Spinal Cord Motor Tracts]

A simple method for estimating global DNA methylation using bisulfite PCR of repetitive DNA elements - http://nar.oxfordjournals.org/cgi/content/full/32/3/e38

Category: Epigenetics and Methylation Protocols: Bisulfite Treatment Methylation Assay Protocol

Yang et al, NAR. 2004. ue to the heavy methylation of repetitive elements, this assay was especially useful in detecting decreases in DNA methylation, and this assay was validated by examining cell lines treated with the methylation inhibitor 5-aza-2'deox - [Read A simple method for estimating global DNA methylation using bisulfite PCR of repetitive DNA elements]

Advances in Cytochemical Methods for Detection of Apoptosis - http://www.jhc.org/cgi/content/full/49/7/821

Category: Cell Biology and Cell Culture: Apoptosis Protocols

Advances in Cytochemical Methods for Detection of Apoptosis. Katherine L. Barrett et al. 2001 - [Read Advances in Cytochemical Methods for Detection of Apoptosis]

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]

Bacteriophage Lamda Plant Genomic Library Construction Protocol - http://www.oardc.ohio-state.edu/stockingerlab/Protocols/GenomicDNAPhageLibraryConstruction.pdf

Category: DNA Protocols: Genomic DNA Library Protocols

Protocol for bacteriophage lamda plant genomic library construction. Includes: Preparation of the insert DNA; Pilot Sau3A Digestion; Full-scale partial digestions. - [Read Bacteriophage Lamda Plant Genomic Library Construction Protocol]

Cap-Switching RACE Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/14/pdb.prot4133

Category: PCR Protocols: cDNA Synthesis Protocols

This method, for the selective amplification of full-length cDNA ends, involves the addition of an adapter during reverse transcription. This method takes advantage of the propensity of Moloney murine leukemia virus reverse transcriptase (MMLV RT) to append two to four cytosines to the 3'-end of newly synthesized cDNA strands. The additional residues are added when the enzyme reaches the 5'-cap structure at the end of the mRNA template. - [Read Cap-Switching RACE Protocol]

Characterization of Protein Glycosylation Using Chip-Based Nanoelectrospray with Precursor Ion Scann - http://jbt.abrf.org/cgi/content/full/16/3/209

Category: Molecular Biology Protocols: Carbohydrate Protocols

Characterization of Protein Glycosylation Using Chip-Based Nanoelectrospray with Precursor Ion Scanning Quadrupole Linear Ion Trap Mass Spectrometry. Sheng Zhang1 and Brian L. Williamson. Journal of Biomolecular Techniques, 16:209-219 2005. - [Read Characterization of Protein Glycosylation Using Chip-Based Nanoelectrospray with Precursor Ion Scann]

Construction of cDNA Libraries Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot3560

Category: DNA Protocols: cDNA Library Protocols

Here, the DNA-RNA hybrids synthesized in Stage 1 are converted into full-length double-stranded cDNAs. The primers for synthesis of second-strand cDNA are created by RNase H, which introduces nicks into the RNA moiety of the cDNA-mRNA hybrids. E. coli DNA polymerase I extends the newly created 3'-hydroxyl termini, using the first-strand cDNA as a template. - [Read Construction of cDNA Libraries Protocol]

Cytochemical Methods for the Detection of Apoptosis - http://www.jhc.org/cgi/content/full/47/9/1101

Category: Cell Biology and Cell Culture: Apoptosis Protocols

Cytochemical Methods for the Detection of Apoptosis. Mark C. Willingham 1999 - [Read Cytochemical Methods for the Detection of Apoptosis]

Extraction and analysis of diagnostically useful proteins from formalin-fixed, paraffin-embedded tis - http://www.jhc.org/cgi/content/full/46/3/397

Category: Protein Protocols: Protein Extraction From Tissue

Extraction and analysis of diagnostically useful proteins from formalin-fixed, paraffin-embedded tissue sections. Ikedaa et al, 1998 JHC. - [Read Extraction and analysis of diagnostically useful proteins from formalin-fixed, paraffin-embedded tis]

Fluorescent indicators for imaging protein phosphorylation in single living cells - http://www.nature.com/nbt/journal/v20/n3/full/nbt0302-287.html

Category: Protein Protocols: Protein Phosphorylation Protocols

Fluorescent indicators for imaging protein phosphorylation in single living cells. Using Phocuses, genetically encoded fluorescent indicators, one can visualize signal transduction based on protein phosphorylation in living cells. Moritoshi Sato1, Takeaki Ozawa1, Kouichi Inukai2, Tomoichiro Asano2 & Yoshio Umezawa1. Nature Biotechnology - [Read Fluorescent indicators for imaging protein phosphorylation in single living cells]

Fractional Precipitation of Proteins by Ammonium Sulfate - Subscription - http://www.cshprotocols.org/cgi/content/full/2006/16/pdb.prot4309

Category: Protein Protocols: Protein Precipitation Procols

Fractional Precipitation of Proteins by Ammonium Sulfate Simpson Cold Spring Harbor Protocols.2006; 2006: pdb.prot4309 - [Read Fractional Precipitation of Proteins by Ammonium Sulfate - Subscription]

High-resolution SNP mapping by denaturing HPLC - http://www.pnas.org/cgi/content/full/99/16/10575

Category: DNA Protocols: SNP Protocols

High-resolution SNP mapping by denaturing HPLC. A SNP mapping procedure that relies on resolving polymorphisms by denaturing HPLC without the necessity of determining the nature of the SNPs. They demonstrate the use of denaturing high-performance liquid chromatography to identify mutations in the candidate genes and to fine-map chromosomal breakpoints. - [Read High-resolution SNP mapping by denaturing HPLC]

How to do a Western Blot - http://www.well.ox.ac.uk/flint/Western.htm

Category: Protein Protocols: Western Blot Protocol

All the products and steps to complete a full western blotting analysis. (Jonathan Flint lab). - [Read How to do a Western Blot]

Improved Detection of Apoptotic Cells in Archival Paraffin Sections: Immunohistochemistry Using Anti - http://www.jhc.org/cgi/content/full/50/4/449

Category: Cell Biology and Cell Culture: Apoptosis Protocols

Improved Detection of Apoptotic Cells in Archival Paraffin Sections: Immunohistochemistry Using Antibodies to Cleaved Caspase 3. Gown et al, 2002 - [Read Improved Detection of Apoptotic Cells in Archival Paraffin Sections: Immunohistochemistry Using Anti]

Isolation of Plant Transcription Factors Using a Modified Yeast One-Hybrid System - http://www.plantmethods.com/content/2/1/3

Category: Plant Biology Protocols: Plant Genetics Protocols

The preparation of expressional cDNA libraries for use in the yeast two-hybrid system is quick and efficient when using the dedicated Clontech™ product, the MATCHMAKER Library Construction and Screening Kit 3. This kit employs SMART technology for the amplification of full-length cDNAs, in combination with cloning using homologous recombination. - [Read Isolation of Plant Transcription Factors Using a Modified Yeast One-Hybrid System]

Laser-capture Microdissection Protocol - http://www.nature.com/nprot/journal/v1/n2/full/nprot.2006.85.html#a1

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

LCM technology can harvest the cells of interest directly or can isolate specific cells by cutting away unwanted cells to give histologically pure enriched cell populations. A variety of downstream applications exist: DNA genotyping and loss-of-heterozygosity (LOH) analysis, etc. Protocol provides a thorough description of LCM techniques, with an emphasis on tips and troubleshooting advice derived from LCM users. The total time required to carry out this protocol is typically 1–1.5 h. - [Read Laser-capture Microdissection Protocol]

Light and Electron Microscopy Detection of Apoptosis - http://www.jhc.org/cgi/content/full/47/4/561

Category: Cell Biology and Cell Culture: Apoptosis Protocols

Detection of Fragmented DNA in Apoptotic Cells Embedded in LR White:Histochemical (LM) and Ultrastructural (EM). Goping et al 1999. - [Read Light and Electron Microscopy Detection of Apoptosis]

Live-Cell Imaging of GFP in Plants - http://www.cshprotocols.org/cgi/content/full/2007/3/pdb.ip31

Category: Microscopy Protocols: Live-Cell Imaging Protocols

GFP serves as a molecular marker that can be imaged dynamically in living cells, both in its native form & as a fusion to other proteins. For GFP imaging, plants present the challenge of autofluorescence from chlorophyll, lignified cell walls, vacuolar contents, and other cell materials, all of which can obscure the GFP signal. Maximizing the signal-to-noise ratio is a major concern, and careful consideration should be given to the choice of tissue imaged, GFP expression level, etc. - [Read Live-Cell Imaging of GFP in Plants]

Methylation profiles of genes utilizing newly developed CpG island methylation microarray on colorec - http://nar.oxfordjournals.org/cgi/content/full/33/5/e46

Category: DNA Microarray Protocols: DNA Methylation Microarray CpG

Kimura et al., NAR 2005. "easy-handling technology provided reproducible and precise measurement of methylated CpGs in MGMT promoter and, thus, our method may bring about a potential evolution in the handling of a variety of high-throughput DNA methylatio - [Read Methylation profiles of genes utilizing newly developed CpG island methylation microarray on colorec]

Microinjection of dsRNA into Drosophila Embryos Protocol - http://www.cshprotocols.org/cgi/content/full/2006/21/pdb.prot4516

Category: Microinjection Protocols: Microinjection of DNA Protocols

This protocol provides a description of how to introduce double-stranded RNA (dsRNA) into Drosophila embryos by microinjection. Several days of preparation are required before injections into Drosophila embryos begin. Flies must be in abundant supply for egg collection. Bombardment of embryos with dsRNA-coated gold particles (Delivery of dsRNA into Drosophila Embryos by a Gene Gun) can be used as an alternative. - [Read Microinjection of dsRNA into Drosophila Embryos Protocol]

Simplifying hESC culture - http://www.bloodjournal.org/cgi/content/full/105/12/4550

Category: Cell Biology and Cell Culture: ES Stem Cell Biology and Culture: ES Cell Growth Media and Requirements

Human embryonic stem cells are a valuable resource for research and cell replacement therapy but are notoriously cumbersome to culture. Bhatia and colleagues show that an increased dose of basic fibroblast growth factor eliminates the need for feeder laye - [Read Simplifying hESC culture]

Transcript In Situ Hybridization of Whole-Mount Embryos for Phenotype Analysis of RNAi-Treated - http://www.cshprotocols.org/cgi/content/full/2006/21/pdb.prot4519

Category: Cytogenetics Protocols: In Situ Hybridization Protocols

This protocol fixes and prepares embryos for in situ hybridization to visualize transcript expression patterns. It is a modification of the method developed by Tautz and Pfeifle for whole-mount in situ analysis of embryos. Use of the standard hybridization protocol on RNAi-treated embryos results in high background staining, which makes visualization of transcript expression patterns practically impossible. The following modifications eliminate this problem and allow visualization of transcript. - [Read Transcript In Situ Hybridization of Whole-Mount Embryos for Phenotype Analysis of RNAi-Treated]

Articles (1-7 of 7)

Single Stranded Plasmid DNA Isolation Protocol

A Single Stranded Plasmid DNA Isolation Protocol describing the production and isolation of single-stranded DNA (ssDNA) using bacteriophagemid-containing bacteria and helper phage. Infection of the host cells with helper phage allows for packaging of ssDNA into bacteriophage. The ssDNA can then be isolated from phage particles.

Probe Preparation for 22 x 40 mm DNA Microarrays

Probe Preparation for 22 x 40 mm DNA Microarrays Protocol.

In Vitro Translated Xenopus Mos Kinase Assay Protocol

In Vitro Translated Xenopus Mos Kinase Assay Protocol. In response to progesterone, immature Xenopus oocytes mature to eggs that can be fertilized. The Mos protein kinase is essential for oocyte maturation, most likely due to its ability to activate the MAP kinase cascade. This MAP kinase cascade eventually leads to the activation of Cdc2/cyclin B and entry into M phase. In this protocol, tagged Mos kinase is translated in vitro, immunopurified, and used in a kinase assay.

DNA Fragment Purification from Polyacrylamide Gels Protocol

A simple protocol for the purification of DNA fragments from Polyacrylamide Gels.

Picking Transfected ES Cell Clones Protocol

This protocol a protocol on how to generate transfected embryonic stem (ES) cell clones. The previous protocol in this series is the Protocol for Electroporation of ES cells. The next protocol in the series is the Protocol on Disaggregation, Expansion, and Freezing of Transfected ES Clones.

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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