New England BioLabs
Monarch® Plasmid DNA Miniprep Kit
NEB #T1010
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The aim of this project was to obtain the full DNA sequence of a gene amplified in our new mouse model. We observed a small change in the molecular weight of this gene compared to wild type mice. Therefore, we isolated the DNA fragment from the agarose gel, used E. coli and plasmid vectors to generate recombinant DNA that was isolated using the Miniprep kit and sent out for sequencing. I used this product because I only needed to do this a few times and did not want to invest in a larger kit. It was inexpensive and came is smaller quantities than the mini-prep kit we used before.
Plamid DNA isolation
E. coli
Resuspend the cell pellet by vortexing in Plasmid Resuspension Buffer, then lyse the cells by adding Plasmid Lysis Buffer. Gently invert tubes about 6 times until color changes to dark pink. Do not vortex. Neutralize the lysate with the Plasmid Neutralization Buffer. Invert tube until color changes to yellow. Centrifuge the tubes and transfer the supernatant to the spin column. Wash the column before eluting the DNA with EB buffer.
All wash steps are required and not optional.
We obtained very high yields of plasmid DNA
N/A
Reliable and inexpensive
None
Great alternative to other miniprep kits.