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Discover the Microbes Within! Wolbachia Project
Приєднався 15 кві 2021
Discover the Microbes Within: The Wolbachia Project is an integrative lab series designed to bring real-world scientific research into middle, high school and college biology classes through inquiry, discovery, biotechnology, and a culture of excellence. Our goals are to engage students in nature and real-world research, and to contribute new scientific data on the bacterial endosymbiont Wolbachia pipientis.
PCR: Program the Thermal Cycler
o This clip shows an example of a thermal cycler being programmed for PCR following the standard protocol for CO1 Arthropod DNA amplification.
o For the full protocol and introduction to PCR, visit: wolbachiaproject.org/pcr/
o To get involved with The Wolbachia Project, visit us at wolbachiaproject.org/
o All of our curriculum is available under the Creative Commons Attribution-Non-Commercial- No Derivatives International License. Contact info(at) wolbachiaproject(dot)org if you would like to make adaptations beyond the classroom.
o For the full protocol and introduction to PCR, visit: wolbachiaproject.org/pcr/
o To get involved with The Wolbachia Project, visit us at wolbachiaproject.org/
o All of our curriculum is available under the Creative Commons Attribution-Non-Commercial- No Derivatives International License. Contact info(at) wolbachiaproject(dot)org if you would like to make adaptations beyond the classroom.
Переглядів: 174
Відео
PCR: Collect and Label a 1.5 ml tube
Переглядів 8762 місяці тому
o This clip shows a sterile 1.5 ml tube being collected and labeled “A” for Arthropod PCR Cocktail. For the Wolbachia PCR cocktail, use the letter “W.” o For the full protocol and introduction to PCR, visit: wolbachiaproject.org/pcr/ o To get involved with The Wolbachia Project, visit us at wolbachiaproject.org/ o All of our curriculum is available under the Creative Commons Attribution-Non-Com...
PCR: Collect and Label 0.2 ml PCR Tubes
Переглядів 8952 місяці тому
o This clip shows six sterile 0.2 ml PCR tubes being collected and labeled 1-6. Make sure to label the top and sides of the tubes and note which Arthropod sample is designated to each tube. When doing the Wolbachia PCR protocol, be sure to use different numbers/names to avoid mixing the samples up. o For the full protocol and introduction to PCR, visit: wolbachiaproject.org/pcr/ o To get involv...
PCR: Add Template DNA to PCR Tubes
Переглядів 1,3 тис.2 місяці тому
o This clip demonstrates template DNA being added to each 0.2 ml PCR tube. It is imperative to change tips between each sample to avoid cross-contamination. o For the full protocol and introduction to PCR, visit: wolbachiaproject.org/pcr/ o To get involved with The Wolbachia Project, visit us at wolbachiaproject.org/ o All of our curriculum is available under the Creative Commons Attribution-No...
PCR: Prepare the PCR Cocktail
Переглядів 1 тис.2 місяці тому
o This clip demonstrates how to calculate the amount of each reagent needed to make a “PCR cocktail,” which contains forward and reverse primers, nuclease-free water, and a Taq polymerase master mix. When making the mixture, make enough for the number of reactions 1 to account for any liquid lost in pipetting. Make sure to change tips between each reagent. o For the full protocol and introducti...
PCR: Vortex the PCR Cocktail
Переглядів 4202 місяці тому
o This clip shows the PCR cocktail being briefly vortexed to properly mix all the reagents. o For the full protocol and introduction to PCR, visit: wolbachiaproject.org/pcr/ o To get involved with The Wolbachia Project, visit us at wolbachiaproject.org/ o All of our curriculum is available under the Creative Commons Attribution-Non-Commercial- No Derivatives International License. Contact info(...
PCR: Add the PCR Cocktail to PCR Tubes
Переглядів 1,6 тис.2 місяці тому
o This clip demonstrates the PCR cocktail being added to each 0.2 ml PCR tube. It is vital to use a fresh pipette tip for each transfer to avoid cross-contamination. o For the full protocol and introduction to PCR, visit: wolbachiaproject.org/pcr/ o To get involved with The Wolbachia Project, visit us at wolbachiaproject.org/ o All of our curriculum is available under the Creative Commons Attri...
PCR: Spin Down PCR Tubes
Переглядів 9992 місяці тому
o This clip shows the 0.2 ml tubes containing the DNA samples and PCR cocktail being briefly centrifuged to collect all liquid at the bottom. Ensure that the lids are fully closed and balance the centrifuge before spinning. o For the full protocol and introduction to PCR, visit: wolbachiaproject.org/pcr/ o To get involved with The Wolbachia Project, visit us at wolbachiaproject.org/ o All of ou...
PCR: Start the PCR Program
Переглядів 6322 місяці тому
o This clip demonstrates samples being loaded into a thermal cycler and the PCR program being started. o For the full protocol and introduction to PCR, visit: wolbachiaproject.org/pcr/ o To get involved with The Wolbachia Project, visit us at wolbachiaproject.org/ o All of our curriculum is available under the Creative Commons Attribution-Non-Commercial- No Derivatives International License. Co...
PCR: Spin Down the PCR Cocktail
Переглядів 3552 місяці тому
o This clip shows the PCR cocktail being briefly centrifuged (3-5 seconds) to collect all liquid at the bottom of the tube. Before spinning, make sure to balance the rotor using another tube with a similar volume of liquid. o For the full protocol and introduction to PCR, visit: wolbachiaproject.org/pcr/ o To get involved with The Wolbachia Project, visit us at wolbachiaproject.org/ o All of ou...
Gel Electrophoresis with the MiniOne System: Visualize the Gel
Переглядів 764 місяці тому
o This clip demonstrates how to visualize the results of gel electrophoresis using the MiniOne Electrophoresis System: theminione.com/electrophoresis-system/. Turn on the bright light and place a camera over the viewing hole to take a photo. o For more information and an introduction to gel electrophoresis, visit: wolbachiaproject.org/laboratory-equipment/ o To get involved with The Wolbachia P...
Gel Electrophoresis Using the MiniOne System: Run the Gel
Переглядів 824 місяці тому
o This clip demonstrates how to run an electric current through an agarose gel using the MiniOne Electrophoresis System. Place the orange hood over the chamber and press the power button to activate the current. A green light should appear, and bubbles will form in the running buffer. Allow the gel to run for around 20 minutes, periodically turning on the dim light to check if any bands are pre...
Gel Electrophoresis Using the MiniOne System: Load the Gel
Переглядів 654 місяці тому
o This clip demonstrates how to load an agarose gel using the MiniOne Electrophoresis System. Using a P20 pipette, carefully load 5 l of the DNA ladder into the first well, followed by 5 l of each sample. Hover the tip directly above the well (inside of the buffer solution) when loading the samples. The dim light can be used to illuminate the wells as they are loaded. Use caution to not punct...
Gel Electrophoresis Using the MiniOne System: Add the Running Buffer
Переглядів 434 місяці тому
o This clip demonstrates how to add the running buffer to the gel electrophoresis chamber using the MiniOne Electrophoresis System. Measure around 140 ml of Buffer TBE and pour it into the chamber up to the fill line, ensuring that the gel is completely submerged. o For more information and an introduction to gel electrophoresis, visit: wolbachiaproject.org/laboratory-equipment/ o To get involv...
Gel Electrophoresis Using the MiniOne System: Assemble the Chamber
Переглядів 504 місяці тому
o This clip demonstrates how to prepare the agarose solution using the MiniOne Electrophoresis System. Gently remove the combs by lifting them straight up and out of the agarose gel and clean any residual agarose off the bottom of the tray. Insert the plastic box into the chamber, ensuring that the wells are aligned with the negative side of the chamber. Insert the gel and tray into the box and...
Gel Electrophoresis Using the MinOne System: Pour the Gel
Переглядів 384 місяці тому
Gel Electrophoresis Using the MinOne System: Pour the Gel
Gel Electrophoresis Using the MiniOne System: Prepare the Casting Tray
Переглядів 384 місяці тому
Gel Electrophoresis Using the MiniOne System: Prepare the Casting Tray
Gel Electrophoresis Using the MiniOne System: Prepare the Agarose Solution
Переглядів 484 місяці тому
Gel Electrophoresis Using the MiniOne System: Prepare the Agarose Solution
DNA Extraction with QIAGEN DNeasy Kit: Prepare and Label the Sample Tubes
Переглядів 1455 місяців тому
DNA Extraction with QIAGEN DNeasy Kit: Prepare and Label the Sample Tubes
DNA Extraction with QIAGEN DNeasy Kit: Add buffer ATL
Переглядів 1845 місяців тому
DNA Extraction with QIAGEN DNeasy Kit: Add buffer ATL
DNA Extraction with QIAGEN DNeasy Kit: Add Proteinase K
Переглядів 3465 місяців тому
DNA Extraction with QIAGEN DNeasy Kit: Add Proteinase K
DNA Extraction with QIAGEN DNeasy Kit: Add Buffer AL
Переглядів 2055 місяців тому
DNA Extraction with QIAGEN DNeasy Kit: Add Buffer AL
DNA Extraction with QIAGEN DNeasy Kit: Incubate at 56°C
Переглядів 4155 місяців тому
DNA Extraction with QIAGEN DNeasy Kit: Incubate at 56°C
DNA Extraction with QIAGEN DNeasy Kit: Centrifuge to Pellet Debris
Переглядів 2755 місяців тому
DNA Extraction with QIAGEN DNeasy Kit: Centrifuge to Pellet Debris
DNA Extraction with QIAGEN DNeasy Kit: Add Ethanol
Переглядів 3355 місяців тому
DNA Extraction with QIAGEN DNeasy Kit: Add Ethanol
DNA Extraction with QIAGEN DNeasy Kit: Prepare and Label Spin Columns
Переглядів 5565 місяців тому
DNA Extraction with QIAGEN DNeasy Kit: Prepare and Label Spin Columns
DNA Extraction with QIAGEN DNeasy Kit: Centrifuge for One Minute
Переглядів 3295 місяців тому
DNA Extraction with QIAGEN DNeasy Kit: Centrifuge for One Minute
DNA Extraction with QIAGEN DNeasy Kit: Wash Step 1
Переглядів 9295 місяців тому
DNA Extraction with QIAGEN DNeasy Kit: Wash Step 1
DNA Extraction with QIAGEN DNeasy Kit: Wash Step 2
Переглядів 6545 місяців тому
DNA Extraction with QIAGEN DNeasy Kit: Wash Step 2
DNA Extraction with QIAGEN DNeasy Kit: Prepare and Label Tubes for Extracted DNA
Переглядів 845 місяців тому
DNA Extraction with QIAGEN DNeasy Kit: Prepare and Label Tubes for Extracted DNA
Why are you not making other videos cDNA, RNA extraction
Great explanation and visuals!
Thankyou so much from Gadjah Mada University ❤
How about i incubate single body mosquito in 3 hours? Is that more effective than ine hour ?
Thankyou so much ❤️
Please I am looking for lab contact work on Wolbachia, I am looking for a postdoc position
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?
i have one question. can we preserve whitefly in 70% Ethanol and extract DNA after 2 months? is it possible?
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.
really Thank you so much for sharing those information, as a debutant in molecular biology, I learned a lot from you
thank you for the informative video, / this is Dr Takwa from Tunisia, recently I defended my phd thesis. during my phd I screened wolbachia in the insect I work on. As a post doc, I am very interested to find an internship in this field of work to build my skills and learn more. could you please help me in that?thank you
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.
This protocol is optimized with a shorter incubation time for classroom use. If you reduce the amount of proteinase K, we recommend increasing the incubation time to enhance lysis. Also make sure to efficiently grind the sample with micropestles.
Hi, your video really helped me! it is well presented short videos, with easy understanding language for science student, and the explanation was really awesome!
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
Romario, its good that u also watched this video.😄
@@saranga6304 🤣🤣🤣🤣
Yes, Wolbachia can be detected in legs as reported here: www.ncbi.nlm.nih.gov/pmc/articles/PMC3342236/. Most of our participants isolate from fresh samples, but some have successfully amplified DNA from from dead organisms. Make sure to indicate the state of your organism along with any reported data.
I wonder how many people disliked
Who else is here for DB’s class -->
Yo waddup HRA squad
Why not make albicans commercially available.
gzzqp vyn.fyi
This is a great introduction. Thanks, Athena!
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?
Great question. Buffer AL is categorized as a lysis buffer, so we added it at 2:47 during the "Cell Lysis and DNA Precipitation" portion of our protocol. Here's a link to the protocol we use, which uses the DNeasy kit for insect extraction: www.qiagen.com/us/resources/resourcedetail?id=cabd47a4-cb5a-4327-b10d-d90b8542421e&lang=en I hope that helps, please let us know if you have further questions! Also feel free to email us at wolbachiaproject(at)vanderbilt(dot)edu.
Great VIdeo! Waiting for Lab 5 ;)
We're hoping bioinformatics whisperer Sarah Bordenstein will feature in Lab 5! :)
This was very helpful!
We love to hear that, thanks!
These videos are great! I'm planning to participate in the Wolbachia Project next year and will definitely use these videos with my students. Thanks!
Thanks, Dan! We're excited to be able to be able to produce these kinds of videos to help students. Let us know if you need anything!