Basket Updated
0
Items added
There are a number of markers that can be used to distinguish the many cell types of the central and peripheral nervous system during development, adult neurogenesis, and the pathogenesis of neurodegenerative disorders.
If a dot blotting apparatus is not available for use, we recommend denaturing your DNA in a lower volume/higher concentration (see modified steps below):Section B. Dot Blot Modified Steps:1. Dilute fragmented genomic DNA to 400 ng/ul in 100 ul of nuclease-free water. Then denature DNA by adding 100 ul of 2X DNA Denaturing Buffer and incubating at 95C for 10 min.2. Add 200 ul of 20X SSC buffer and immediately chill on ice for 5 min. DNA concentration of the solution is 100 ng/ul.3. Set up a series of six 2-fold dilutions by adding 200 ul of the DNA solution, starting with the DNA solution in Step 2, to 200 ul of nuclease-free water. This will generate seven DNA samples containing 200 ul DNA at concentrations of 100 ng/ul, 50 ng/ul, 25 ng/ul, 12.5 ng/ul, 6.25 ng/ul, 3.125 ng/ul and 1.563 ng/ul.4. Spot 10 ul of each of the seven dilution samples onto the nylon membrane, leaving the last well for nuclease-free water only. The amount of DNA added to each well should then be 1000 ng, 500 ng, …
Learn DNA library preparation for ChIP-seq and CUT&RUN using the Illumina® protocol, from end repair & adaptor ligation to PCR enrichment & cleanup.
ChIP-seq is a powerful technique that combines ChIP and next-generation sequencing (NGS) to study DNA-protein interactions across the entire genome, but it is only as good as the quality of the antibody used in the ChIP experiment.
CUT&Tag delivers quality NGS data even if the signal from the Bioanalyzer or TapeStation System is low and the DNA library yield is low.
In the CUT&RUN kit protocol, the addition of digitonin to the buffers facilitates the permeabilization of cell membranes & the entry of the primary antibody & pAG-MNase enzyme into the cells & nuclei.
Apoptosis is a type of programmed cell death that occurs normally throughout the lifespan and can also occur in response to harmful stimuli. Apoptotic cells are identified by their altered morphology, caspase activation, and the presence of damaged DNA.
Multiplex Oligos for Illumina Protocol (Single Index Primers) (ChIP-seq, CUT&RUN): easy to follow directions describing the step by step experimental procedure.a
CUT&RUN Frequently Asked Questions (FAQs) from Cell Signaling Technology, Inc.
Case Study 4: Exceptional Performance of XP Monoclonal Antibodies from Cell Signaling Technology.
The typical DNA yields from ChIP experiments range from 0.2-2 ng/µl when targeting transcription factors or cofactors and from 2-20 ng/µl when targeting histone modifications. If the concentration is within this range, then you should be able to continue with ChIP-seq. Our DNA Library Prep Kit for Illumina® (ChIP-seq, CUT&RUN) #56795 can generate a library with 0.5ng ChIP-DNA.
Discover the key epigenetic drivers of breast cancer and the best products to target them.
Multiplex Oligos for Illumina Protocol (Dual Index Primers) (ChIP-seq, CUT&RUN): easy to follow directions describing the step by step experimental procedure.
Fostering mentorship, application-based learning, and a love of science are all part of the mission of the CST internship program. Students interning with us are partnered with CST Mentors.
Discover the most important epigenetic markers that contribute to the progression of diffuse intrinsic pontine glioma (DIPG).
Cellular senescence is characterized by irreversible cell-cycle arrest in combination with a distinct secretory phenotype and expanded lysosomes in response to stress. Senescent cells accumulate in tissues during aging, and various markers of senescence
Discover the top epigenetic markers that you should look out for in prostate cancer research.
Chromatin Immunoprecipitation Workflow Solutions for ChIP-qPCR and ChIP-seq
If you start with 100,000 cells, you typically get 0.6-6ng DNA. However, if you start with 5,000 - 10,000 cells, the DNA yield can be as low as 0.1-0.2 ng or even undetectable by the picogreen assay. In this case, we suggest you directly proceed with library prep using 20 PCR amplification cycles, which will result in a library DNA with a concentration of around 10-30 ng/ul. QC of the CUT&RUN DNA by qPCR can be performed on the library DNA if you hope to save as much CUT&RUN DNA as possible for the library prep.
WB bands not where they're supposed to be? In addition to mAb troubleshooting, you can visualize the total protein by staining with Ponceau or Coomassie.
Usually, the CUT&Tag DNA libraries from histone targets have a higher concentration than those from non-histone targets. We use the following formula to convert a library concentration from ng/µL to nM before diluting each library sample to the same concentration (nM) for pooling purposes: Concentration (nM) = 1,000,000 X Concentration (ng/µL) / library average size (bp) / 660. In addition to obtaining a greater library size for undetectable library samples from the Bioanalyzer or TapeStation system (as described in the above question), we would also suggest pooling the libraries that have a flat signal of 5-10 fold more than the libraries that show normal sized peaks on the Bioanalyzer or TapeStation systems. This ensures an even distribution of the number of reads among all samples. Usually, a library pool concentration of 2 nM DNA is enough for NGS purposes, although a higher concentration is always welcome.
CUT&Tag Frequently Asked Questions (FAQs) from Cell Signaling Technology, Inc.
Cell viability assays measure the population of live, viable cells within a sample. Typically, viability assays measure markers of cell health, including cellular metabolism, ATP levels, and cell proliferation.
Case Study 2: Exceptional Sensitivity of XP Monoclonal Antibodies pertaining to eXceptional Monoclonal Technology (XMT) from Cell Signaling Technology.
Find out which epigenetic markers are important drivers in mixd lineage leukemia, or MLL.
ELISA-Peptide Assay Protocol : easy to follow directions describing the step by step experimental procedure.
