Behind the Bench: DNA Extraction with Qiagen DNeasy Blood & Tissue Kit (Product 69504)

In Lab 2 of The Wolbachia Project, we will be extracting DNA from your arthropod sample using Qiagen’s DNeasy Blood and Tissue Kit (Product 69504). This timelapse video will show you the steps of this protocol behind the bench, and The Wolbachia Project Database.

0:00 Introduction
0:14 Prepare Materials and Workspace
1:02 How to Dissect an Arthropod
1:52 Sample Preparation
2:14 Cell Lysis & DNA Precipitation
3:43 DNA Purification
6:18 DNA Elution
7:12 How to Load the Centrifuge for Elution
7:46 Finish DNA Elution
8:16 Clean Up
8:33 The Wolbachia Project Database
8:47 Outro

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[ccdj]

3:37 You only mentioned ethanol here. The protocol I have suggests an ethanol(90ml)-AL(86ml) mixture. Thanks for the video though. For someone like me who never worked in a lab, it was very useful.

[juanayala]

Thanks for video! It has helped me a lot. I just have one question. I noticed that in the DNA precipitation step, you didn't use the AL buffer as is described in the manual of the product. So, is that buffer optional or just for specific samples?

[beckycloud]

My lab is worried about splash risk while working with field-collected mosquitoes that may contain human pathogens, so pestle homogenization is not ideal for me. Would 3mm stainless steel beads work fine?

[itsjatt]

i have one question. can we preserve whitefly in 70% Ethanol and extract DNA after 2 months? is it possible?

[anastasiating]

Can we extract the DNA from the legs of insects? And do we need to get a fresh sample or alive insects for DNA extraction

[tasnimbintaosman]

We Didn’t get dna. All lines we followed except 200 protinase K. We used 30 protinase K. Among 8 samples, we got for two.