HiPurA™ Streptomyces DNA Purification Kit

Protocol tips

Upstream tips	Protocol tips	Downstream tips
Examine the reagents for precipitation. If any kit reagent forms a precipitate (other than enzymes), warm at 55-65°C until the precipitate dissolves and allow cooling to room temperature (15-25°C) before use.

Upstream tips
Examine the reagents for precipitation. If any kit reagent forms a precipitate (other than enzymes), warm at 55-65°C until the precipitate dissolves and allow cooling to room temperature (15-25°C) before use.

Publication protocol

Genomic DNA isolation
The genomic DNA was isolated using the Hipura Streptomyces DNA spin kit-MB 527-20 pr (Himedia), according to the manufacturer's protocol. Briefly, the freshly cultured cells were pelleted by centrifuging for 2 minutes at 10,000g to obtain 10–15 mg (wet weight). The cells were resuspended thoroughly in 300 μL of lysis solution and 20 μL of RNase A solution was added, mixed, and incubated for 2 minutes at room temperature. About 20 μL of the proteinase K solution (20 mg/mL) was added to the sample and mixed; the resuspended cells were transferred to a Hibead Tube and incubated for 30 minutes at 55°C. The mixture was vortexed for 5–7 minutes and incubated for 10 minutes at 95°C followed by pulse vortexing. Supernatant was collected by centrifuging the tube at 8000g for 1 minute at room temperature. About 200 μL of lysis solution was added, mixed thoroughly by vortexing, and incubated at 55°C for 10 minutes. An amount of 200 μL of ethanol (96–100%) was added to the lysate and mixed thoroughly by vortexing for 15 seconds. The lysate was transferred to a new spin column and 500 μL of prewash solution was added to the spin column and centrifuged at 8000g for 1 minute; the supernatant was discarded. The lysate was then washed in 500 μL of wash solution and centrifuged at 8000g for 3 minutes. The elution buffer (200 μL) was pipetted out and added directly into the column without spilling and incubated for 1 minute at room temperature. Finally, the DNA was eluted by centrifuging the column at 8000g for 1 minute.

Full paper   Login or join for free to view the full paper.

Reviews

Discussion

Papers

Check out relevant papers found by Labettor's AI that are relevant for performing DNA isolation / purification Bacteria - Gram positive Streptomyces. Sp using HiPurA™ Streptomyces DNA Purification Kit from HiMEDIA.

Manufacturer protocol

Download the product protocol from HiMEDIA for HiPurA™ Streptomyces DNA Purification Kit below.

Download manufacturer protocol

Videos

Check out videos that might be relevant for performing DNA isolation / purification Bacteria - Gram positive Streptomyces. Sp using HiPurA™ Streptomyces DNA Purification Kit from HiMEDIA. Please note that these videos are representative and steps or experiment specific processes must be kept in mind to expect desired results.