AllPrep DNA/RNA Mini Kit

DNA isolation / purification Tissue - fecal sample

4/5 - 1 Review - 100% recommends this

Experiment
DNA isolation / purification Tissue - fecal sample
Product
AllPrep DNA/RNA Mini Kit from Qiagen
Manufacturer
Qiagen

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

Publication protocol

Colonic mucosal biopsies, 2–3 mm in diameter, were collected from two patients undergoing planned colectomy at Akershus University Hospital, and from three patients undergoing colonoscopy as part of the EU-project IBD-Character (EU ref no 305676). Prior to the colectomy and colonoscopy the patients performed a bowel cleansing by Picoprep (Ferring Legemidler AS, Oslo, Norway) according to manufacturer’s instructions. All biopsies were immediately placed in Allprotect Tissue Reagent (Qiagen, Hilden, Germany) and stored according to manufacturer’s instructions. For each of the different protocols tested, one to two biopsies from each patient were used due to a limited number of biopsies (Table 1).

Total RNA and DNA were purified from colonic mucosal tissue using the AllPrep DNA/RNA Mini Kit (Qiagen). Manufacturer’s instructions were followed with the exception of the lysis steps, where three tissue lysis protocols (Protocol 1, 2 and 3) were performed and evaluated (Fig. 1; Additional file 1). For RNA purification, DNase treatment was included and performed on column as described in the AllPrep DNA/RNA Mini Kit protocol. RNA and DNA were eluted using 40 µl nuclease free water (NFW) and stored at −80 and −20 °C, respectively.

The resulting RNA quantity and quality data from the tested protocols were compared to RNA quantity and quality data obtained using a standard RNA purification protocol combining Qiazol, phase separation and kit column based purification [16] (Table 1; Additional file 1). The resulting DNA quantity and quality data were compared to DNA quantity and quality data from a combination of mechanical and enzymatic pre-treatments as recommended by Qiagen for lysis of Gram-positive bacteria, followed by purificaton with AllPrep DNA/RNA Mini Kit (kit 1), the QIAamp DNA Stool Mini Kit (kit 2) and the DNeasy Blood & Tissue Kit (kit 3) (Table 1; Additional file 1). For all three purification kits the manufactures instructions were followed after pre-treatment. DNA was eluted using 40 µl NFW and stored at −20 °C.

The concentration of the RNA and DNA samples were assessed using NanoDrop ND-1000 spectrophotometer (Thermo Fisher Scientific, Waltham, MA, USA). For RNA quality the RNA integrity number (RIN) was tested using the Agilent 2100 Bioanalyzer (Agilent Technologies Inc., Santa Clara, CA, USA) and the 2100 Expert Software. The assay and reagent kit used were Eukaryote Total RNA Nano Series II and the Agilent RNA 6000 Nano Kit. For DNA the quality was obtained using the ratio 260/280 for assessing the purity of the samples. The molecular size of the genomic DNA was measured in a 1 % agarose gel in 1xTBE, 70 V, running time 2.5 h (Fig. 2).

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

Reviews

Scientist
Luud Roorda
Review date

Protocol
Adjusted
Protocol tips
In use with 0.1 mm zirconia/silica glass beads and Precellys 24 Bertin before putting the lysate into a QIAschredder.
Protocol adjustment
Upstream

Product rating
Would recommend?
Yes

What worked well?
  • high concentrations of DNA
What could be improved?
  • Long protocol
  • not user friendly

Rate review   Login or join for free to view the full paper.

Discussion

Discussion

2 years ago

Author: Denmark

What DNA isolation kit would work for insect samples?

Hello everyone! I am currently using different DNA isolation kits to extract DNA from insects. Even though I am able to successfully extract DNA I would like to maximize the yield. Do you have any tips that might help me with that even if the kits are not specifically designed for insect samples?

Share your thoughts or question with experts in your field by adding a discussion!

Papers

Check out relevant papers found by Labettor's AI that are relevant for performing DNA isolation / purification Tissue - fecal sample using AllPrep DNA/RNA Mini Kit from Qiagen.

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

Manufacturer protocol

Download the product protocol from Qiagen for AllPrep DNA/RNA Mini Kit below.

Download PDF Download manufacturer protocol

Videos

Check out videos that might be relevant for performing DNA isolation / purification Tissue - fecal sample using AllPrep DNA/RNA Mini 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.

We haven't found any additional videos for this experiment / product combination yet.

Become shareholder Discussions About us Contact Privacy Terms