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CRISPR Mouse

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Get tips on using CMV-CAS9-2A-GFP Plasmid to perform CRISPR Human - Deletion TRIB1

Products Sigma-Aldrich CMV-CAS9-2A-GFP Plasmid

Get tips on using pSpCas9(BB)-2A-Puro (PX459) to perform CRISPR Human - Deletion NOX4

Products Addgene pSpCas9(BB)-2A-Puro (PX459)

Cell culture media 3D Cell Culture Media Mouse primary breast cancer ephitelial cells-Mammospheres

The estimation of DNA methylation level heavily depends on the complete conversion of non-methylated DNA cytosines. It is crucial to ensure complete conversion of non-methylated cytosines in DNA. Therefore, it is important to incorporate controls for bisulfite reactions, as well as to pay attention to the appearance of cytosines in non-CpG sites after sequencing, which is an indicator of incomplete conversion.

DNA DNA methylation profiling Gene specific profiling Mouse muscle stem cells SPRY1

The estimation of DNA methylation level heavily depends on the complete conversion of non-methylated DNA cytosines. It is crucial to ensure complete conversion of non-methylated cytosines in DNA. Therefore, it is important to incorporate controls for bisulfite reactions, as well as to pay attention to the appearance of cytosines in non-CpG sites after sequencing, which is an indicator of incomplete conversion.

DNA DNA methylation profiling Whole genome profiling mouse T-cell (CD4 / CD8)

When extracting nucleic acids from cell cultures, thorough homogenization of cells via vortexing in lysis buffer is very necessary. Choose the best RNA isolation method keeping in mind the downstream applications, generally, column-based isolations result in clean and concentrated RNA samples. Downstream applications like sequencing and cDNA synthesis require high-quality RNA, always treat the samples with DNases and check their integrity by running a gel.

RNA RNA isolation / purification Cells primary mouse pulmonary artery smooth muscle cells

Site-directed mutagenesis (SDM) can be challenging, particularly during detection/confirmation of (SDM) in colonies by sequencing or PCR techniques. This common issue in SDM is heavily relying on designing of mutagenic primer pairs. The best solution is to design the mutagenic primers that have extended 3'-ends/3'-overhang. This would provide the annealing region between the mutagenic primer pair is essentially shorter. and hence ensure a lower annealing temperature for the primer pair along with a higher chance of annealing to the template.

DNA Site Directed Mutagenesis (SDM) Mouse Point mutation L929 SigmaR1 gene (σ1)

ELISA is the most commonly used method of detecting and quantifying the concentration of an antigen in an unknown sample. During the experiment, If you get a weak signal, then make sure reagents are at room temperature before starting the assay. Try increasing incubation times to ensure maximal antibody binding and amplify the signal. Secondly, if you get values above 0 in the negative control indicates a high background signal. Try to consider reducing your antibody concentration and prevent non-specific binding of antibodies by using affinity-purified antibody and suitable blocking buffers. To avoid high well to well variation, do not stack plates during incubation, no bubbles in the plate and wash wells thoroughly to avoid variation.

Proteins ELISA Mouse Chitinase-3-Like Protein-1 (CHI3L1) or YKL-40

Get tips on using Monoclonal Anti-MAP Kinase, Activated/monophosphorylated (Phosphothreonine ERK-1&2) antibody produced in mouse to perform Western blotting ERK

Products Sigma-Aldrich Monoclonal Anti-MAP Kinase, Activated/monophosphorylated (Phosphothreonine ERK-1&2) antibody produced in mouse

RNA siRNA / miRNA gene silencing Mouse Glomerular mesangial cells HIPK2 Polymer / Lipid delivery

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