QIAamp Fast DNA Tissue Kit

Protocol tips

Upstream tips	Protocol tips	Downstream tips
		- Include RNAse treatment for 15-20 min. - Ensure EtOH is completely evaporated off of the column prior to elution. Adjust time from 1min to 5 min at 60`C - Use prewarmed TE buffer to elute the DNA

Downstream tips
- Include RNAse treatment for 15-20 min. - Ensure EtOH is completely evaporated off of the column prior to elution. Adjust time from 1min to 5 min at 60`C - Use prewarmed TE buffer to elute the DNA

Publication protocol

2. Quantitative real-time polymerase chain reaction to measure DNA methylation (MethyLight)
Formalin-fixed, paraffin-embedded tissues were used for DNA extraction. Ten sections of 10-μm thick paraffin-embedded tissues were used for DNA extraction. One gastrointestinal pathologist (K.M.K.) reviewed all slides of the dissected tissues and DNA extraction was used the slides containing only cancer tissues. The paraffin was removed from the tissue by rinsing in xylene, and the genomic DNA was isolated using a QIAamp tissue kit (Qiagen, Valencia, CA, USA). Bisulfite conversion was performed using the Zymo EZ DNA methylation kit (Zymo Research Co., Orange, CA, USA) according to the manufacturer’s protocol. After bisulfite conversion of the same amount of the DNA solution, an -based MethyLight control reaction was performed to quantify the number of input target DNA, and the threshold cycle values were confirmed to be comparable between DNA samples. A MethyLight assay to measure the DNA methylation of seven individual genes was performed as reported previously., Supplementary Table 1 lists the primers and probes specific to methylated DNA and used for the MethyLight reactions. The percentage of the methylated reference (PMR) at a specific gene was calculated by dividing the ratio of a sample by the ratio of fully methylated placental DNA and multiplying by 100. The DNA methylation frequencies (PMR >10) were determined for each CpG island locus. CIMP-high was defined as ≥3/5 methylated markers using the five-marker panel, and CIMP-low and CIMP-negative as ≤2/5 methylated markers. Dr Laird’s five panels (CACNA1G, IGF2, NEUROG1, RUNX3, and SOCS1) for the CIMP marker panel were used.



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 Tissue - colon using QIAamp Fast DNA Tissue Kit from Qiagen.

Manufacturer protocol

Download the product protocol from Qiagen for QIAamp Fast DNA Tissue Kit below.

Download manufacturer protocol

Videos

Check out videos that might be relevant for performing DNA isolation / purification Tissue - colon using QIAamp Fast DNA Tissue Kit from Qiagen. Please note that these videos are representative and steps or experiment specific processes must be kept in mind to expect desired results.