QIAamp DNA Mini Kit

DNA isolation / purification Bacteria - Gram positive Clostridium difficile

Experiment
DNA isolation / purification Bacteria - Gram positive Clostridium difficile
Product
QIAamp DNA 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

The gDNA from C. difficile was first extracted using each of the three existing protocols in the QIAamp handbook (11), namely (i) the Gram-positive bacteria (GP) protocol, (ii) the bacterial plate cultures (PC) protocol, and (iii) the bacterial suspension cultures (SC) protocol. For the GP and SC protocols, bacterial density in Trypticase soy broth (TSB) was adjusted to a 0.5 to 1 McFarland standard, using colonies from brucella blood agar, and incubated overnight at 35°C under anaerobic conditions. For the PC protocol, a single colony was suspended in 180 μl of ATL buffer and vortexed. As shown in Fig. 1A, no distinct bands were observed with the GP protocol. A smear was seen on the gel, suggesting shearing of the DNA. Similarly, no gDNA was obtained using the PC protocol (data not shown). Isolation of gDNA using the SC protocol yielded a very faint large-molecular-weight band. However, significant smearing was present (Fig. 1B), and the amount of gDNA obtained was low and variable, ranging from 0.5 to 2.5 μg.



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Discussion

Discussion

4 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

4 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

Check out relevant papers found by Labettor's AI that are relevant for performing DNA isolation / purification Bacteria - Gram positive Clostridium difficile using QIAamp DNA Mini Kit from Qiagen.

Paper title
Optimized Protocol for Simple Extraction of High-Quality Genomic DNA from for Whole-Genome Sequencing
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Manufacturer protocol

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

Download PDF Download manufacturer protocol

Videos

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