Product Description: Price
Isolation of high-quality plasmid DNA from the gemisch of the E. coli cell lysate paves the way for many laboratory as well as clinical genetic applications, among which are clone screening, DNA sequencing, DNA transfection, probe preparation, PCR analysis, and gene therapy, to name but a few. Small-scale plasmid purification (mini-prep) is often used to facilitate clone screening and DNA sequencing. We have modified and streamlined the alkaline lysis mini-prep method of Sambrook et al. (Molecular Cloning, CSH, NY), eliminating the needs for additional RNase treatment and phenol-chloroform extraction. Plasmid DNA can be easily and rapidly purified from an overnight E. coli culture in 15 min with a typical yield of 1 – 5 ug DNA per ml of culture. The DNA is suitable for restriction digestion, probe preparation, and PCR analysis. DNA to be used for sequencing analysis is subjected to an additional 10-min step of PEG precipitation to remove any trace amount of RNA. High-quality DNA sequencing ladders can be easily obtained (almost 1,000 bases resolved per sequencing run) if the plasmid DNA is isolated from a suitable E. coli host strain. The kit (sufficient for 200 mini-preps) is stable for many years if handled and stored properly.
#plasmid #DNA #sequencing #bacteria #DNAcloning #molecularbiology
Kit Components:
Plasmid Solution-1: 20 ml, store at 4ºC (for 200 mini-preps)
Plasmid Solution-2: 20 ml, store at room temperature
Plasmid Solution-3: 20 ml, store at 4ºC
Plasmid Solution-4: 5 ml, store at 4ºC
SDS:
Tris, EDTA, SDS, NaOH, PEG-8000, potasium acetate, acetic acid
Citation:
Role and differential expression of calpastatin mRNA and protein in cultured cardiomyocytes exposed to hypoxic stress
Mol Cell Biochemistry 265:63-70, 2004