Transformation of E. coli with Plasmid DNA - Edvotek Video Tutorial
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- Опубликовано: 7 сен 2024
- For more information on transformation, check out our Quick Guide!
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In the laboratory, scientists can force bacteria like E. coli to acquire exogenous DNA from the surrounding environment through a process called transformation. The newly acquired genetic information is both stable and heritable.
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It was entertaining to watch the experiment along with that funky groove
I would have preferred it without the funky groove. I found it distracting
Sukeshj, you should contact the manufacturer of the cell strains for specific information. This tutorial was designed to complement our transformation kits designed for high school/college teachers.
I did this when I was 13 it was a very fun process
We love to hear that this biotechnology experience was memorable to you!
Love the video! So helpful! Thanks!
swing and a miss 1:19
lmao
thank you for the video and links!
oh my god this video is so good for beginners
Thank you SO MUCH
thank you for the video and links! i am in amoud university somalia
very helpful video. Thank you
What is the function of the Calcium Chloride? Thank you!
The Calcium Chloride helps the DNA stick to the cells. That way, when the cells are heat shocked, the DNA is more easily taken up by the bacterial cells.
calcium chloride increases cell competency or the cell's ability to pick up a plasmid
nice and easy to follow. Thanks. If I understand very well, this is "competent" and "transformation" combined?
Thanks for your feedback!
The cells are made competent when they are resuspended in the calcium chloride solution. The addition of the plasmid DNA and the subsequent heat shock transforms the cells.
what molarity is the calcium chloride solution used ?
How the hell the plasmids are getting though the bacteria cells membrane just because of the heat ? can it get schematised or something ? I'm curious.
It's believed that the heat shock (the change from cold to hot to cold again) makes the bacterial cell wall/membrane more permeable, allowing the DNA molecules to pass into the cell.
is it not necessary to make the cells competent through centrifuging?
We find that centrifuging is only necessary when you wish to store the competent cells at -80C in a glycerol/calcium chloride solution. In this experiment, the competent cells are used right away.
best
Sir/Mam I have two genes 1) 654bp and 2) 294 bp which are optimized for codon biasing and ligated with pET14b. When I am transforming separately to then in BL21 or Rosetta, i am not getting expression, so can help me to understand problem, so I can proceed ????
What was your independent variable and statistical method for analysis
i want to know if Crispr9 made this procedure obsolete
Crispr is an exciting technology that is proving itself to be very versatile. We do believe that bacterial transformation will always be a useful technique in the biotechnology laboratory.
How to get DNA for human Interlukin 7
Excelent video! By the way, what is the gene that encodes human insulin for type 1 diabetes?
The NIH has a great database that describes genes that have been characterized. Here is the info for insulin www.ncbi.nlm.nih.gov/gene/3630 But, remember that the insulin protein goes through several post translational modifications before it is active!
I can now say I watched an instructional video about how to modify E. coli.
Rock on!
what is the reason that put dna in cold ice ? i believe it is to lower the move of cell membrane..but.....exact reason is for heat shock?? AND!!! i can't understand about time you using....15 min...30 min... is it related with principle of experiment???
The exact mechanism through which heat shock works is not entirely known, but it is believed that the combination of the change in temperature plus the chemical solution open pores in the cell wall and membrane through which the DNA passes into the cell. For recovery, we recommend no less than 5 minutes and no more than 30. You will need to refer to your experimental protocol for specifics, this is just a guide.
@@EdvotekInc thanks for teaching! I'm korean high school student and I am impressed by your kindness/// thank you!
Isn't it a conjugation method?
not a transformation?
In nature, DNA is transferred between bacteria using two main methods- transformation and conjugation. In transformation, a bacterium takes up exogenous DNA from the surrounding environment. In contrast, conjugation relies upon direct contact between two bacterial cells. A piece of DNA is copied in one cell (the donor) and then is transferred into the other (recipient) cell. In both cases, the bacteria have acquired new genetic information that is both stable and heritable.
In this case, we are performing a transformation, where we force free plasmid DNA into a bacterial cell using a temperature change and calcium chloride.
Hi, where do you get all this gear and components?
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you guys need to use less distracting music... or none at all. :\
Thanks for the feedback!
1:19 shit
👌👌👌👌