What is pcDNA3 plasmid?
The pcDNA3-DBD plasmid is used to express GAL4 DNA binding domain (DBD, aa1-147) in eukaryotic system.
How do you pick restriction enzymes?
When selecting restriction enzymes, you want to choose enzymes that:
- Flank your insert, but do not cut within your insert.
- Are in the desired location in your recipient plasmid (usually in the Multiple Cloning Site (MCS)), but do not cut elsewhere on the plasmid.
What is the CMV promoter?
The CMV promoter is a commonly used promoter for the production of high level recombinant protein in mammalian cells17. However, the expression level of the transgene driven by CMV promoter decreases with extended culture times because of transcriptional silencing, which is associated with DNA methylation18, 19.
What is mammalian expression?
Mammalian expression is the system of choice for studying the function of a particular protein in the most physiologically relevant environment, because it allows for the highest level of posttranslational processing and functional activity of the protein.
What is pcDNA vector?
This pcDNA™3.1(-) vector is designed for high-level, constitutive expression in a variety of mammalian cell lines. It contains a Geneticin™ selectable marker and a reverse-orientation multiple cloning site.
What happens if you add too much restriction enzyme?
Incomplete digestion is a frequently encountered issue when using restriction endonucleases. Incomplete digestion may occur when too much or too little enzyme is used. The presence of contaminants in the DNA sample can inhibit the enzymes, also resulting in incomplete digestion.
Why are restriction enzymes put in last?
The restriction enzyme is usually the last component added to a reaction to ensure that it is not exposed to extreme conditions. When many similar digests are being prepared, it may be convenient to create premixes of common reagents.
How is CMV promoter activated?
CMV promoter is repressed by p53 and activated by JNK pathway.
Does pcdna contain hygromycin?
It contains a Hygromycin selectable marker and a forward-orientation multiple cloning site. Three untagged versions of pcDNA™3.1 (available separately), each with a different selectable marker (Geneticin®, Zeocin™, or Hygromycin), are for use alone or in co-transfections.
What is the storage temperature of the pcdna™3/hygro (+) vector?
20 µg of this pcDNA™3.1/Hygro (+) vector, as well as an expression control, are supplied supercoiled and lyophilized. Store at -20°C.
What is the hygro (+) expression vector?
This pcDNA™3.1/Hygro (+) vector is designed for high-level, constitutive expression in a variety of mammalian cell lines. It contains a Hygromycin selectable marker and a forward-orientation multiple cloning site. The pcDNA™3.1 Expression Vector Family
What are the features of pcdna™3?
It contains a Hygromycin selectable marker and a forward-orientation multiple cloning site. Three untagged versions of pcDNA™3.1 (available separately), each with a different selectable marker (Geneticin®, Zeocin™, or Hygromycin), are for use alone or in co-transfections. All three vectors offer the following features: For Research Use Only.