NucleoSpin® Plasmid

DNA isolation / purification Bacteria - Gram negative E.coli

Experiment
DNA isolation / purification Bacteria - Gram negative E.coli
Product
NucleoSpin® Plasmid from Macherey Nagel
Manufacturer
Macherey Nagel

Protocol tips

Upstream tips
Buffers A3 and AW contain guanidine hydrochloride which can form highly reactive compounds when combined with bleach (sodium hypochlorite). so DO NOT add bleach or acidic solutions directly to the sample preparation waste.

Publication protocol

Extracted DNA was visualized on a 1% agarose gel stained with GelRed and run at 70 V for 60 min. The DNA was electroporated at 1.8 kV into E. coli DH5α, selected on ampicillin (32 mg/L), tetracycline (16 mg/L), kanamycin (25 mg/L), colistin (16 mg/L), and ciprofloxacin (4 mg/L), or incubated at 37°C overnight. Plasmid DNA was extracted from the transformants using the Machery-Nagel NucleoSpin Plasmid kit and digested with EcoRI restriction enzyme according to the manufacturer's guidelines. Plasmids were visualized on a 1% agarose gel stained with GelRed. Antibiotic susceptibility testing via the disk diffusion method was carried out on transformants according to the CLSI guidelines (Clinical Laboratory Standards Institute, 2016).

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Reviews

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Discussion

Discussion

2 years ago

Author: Israel

How can I improve my DNA yield?

The DNA concentration after using this DNA isolation kit is sometimes too low and thus it is not sufficient for my follow-up experiments. How can I improve it?

Discussion

2 years ago

Author: Milena Alexeyeva Russian Federation

Tips on storing DNA templates?

Hello there! I just started doing experiments on bacterial DNA and I would like your opinion on storing DNA templates. Which are the desired and most optimal conditions?

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Papers

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Manufacturer protocol

Download the product protocol from Macherey Nagel for NucleoSpin® Plasmid below.

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