DNA Isolation Kit for Cells and Tissues

DNA isolation / purification Cells - primary cells human primary keratinocytes

Experiment
DNA isolation / purification Cells - primary cells human primary keratinocytes
Product
DNA Isolation Kit for Cells and Tissues from Sigma-Aldrich
Manufacturer
Sigma-Aldrich

Protocol tips

Protocol tips
During protein precipitation, Samples must be centrifuged at 26,900 × g (rmax) for a minimum of 20 minutes. Lower centrifuge temperatures or speeds will result in very loose protein pellets, reduced yields and contamination of the DNA sample with protein.

Publication protocol

Genomic DNA isolation and sonication
Keratinocytes were washed and trypsinized. Genomic DNA was purified using DNA Isolation kit for cells and tissues (Roche Applied Science). Cells were counted and resuspended in PBS to wash the cells and centrifuged at 1500 rpm for 10 min. Cellular lysis buffer was added and homogenized to a fine suspension. Proteinase K was added and incubated at 65°C for 2 h. Further RNase solution was added and incubated at 37°C for 15 min. Proteins were precipitated, and the supernatant was separated. The genomic DNA was precipitated by using isopropanol. The isolated genomic DNA was diluted and sonicated into small fragments using Sonifier B-12 (Branson Sonic Power, Dietzenbach, Germany). Sonication produced a concoction of random small fragments of DNA of varied lengths.

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Discussion

Discussion

1 year ago

Author: R. Verma India

DNA isolation column clogged

During centrifugation, the column got clogged and I was unable to continue with the protocol. How can I unclog it?

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Papers

Check out relevant papers found by Labettor's AI that are relevant for performing DNA isolation / purification Cells - primary cells human primary keratinocytes using DNA Isolation Kit for Cells and Tissues from Sigma-Aldrich.

Paper title
Inflammatory cytokines break down intrinsic immunological tolerance of human primary keratinocytes to cytosolic DNA.
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Manufacturer protocol

Download the product protocol from Sigma-Aldrich for DNA Isolation Kit for Cells and Tissues below.

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