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Calvin P

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  1. Hello everyone I have a genetics related question on how to achieve a transformation in agrobacterium without having the gene being expressed bu the agrobacterium. According to what I have learned it seems relatively simple to achieve a transient transfection with plasmid vectors and cationic liposomes in bacteria. However my concern is that this approach will result in the agrobacterium expressing the gene instead of adding the gene to it's Ti plasmid. The goal is to insert a firefly luciferase gene into Arabidopsis.
  2. Hello everyone I'd like to learn more about calli and tissue culture in general, my goal is to grow an arabidopsis callus so that I can hopefully insert a gene into it. If anybody has resources related to growing arabidopsis calli please let me know. Additionally if you guys have recommendations for suitable growth medium also let me know. Thank you
  3. Much appreciated @StringJunky by any chance would you know any ebooks or papers I can read to improve my knowledge on this?
  4. I'm currently a student learning about genetic engineering and one thing that I didn't quite understand is before extracting an egg cell from a female, the female mouse must first perform coitus with a vasectomized male mouse. Does anybody know why or how this is related to the female mouse releasing an egg cell?
  5. @Dagl1 Thank you very much for the insights they we're all incredibly helpful, however I still have a few more questions. 1. What's M2 and how do you "wash" egg cells in them? 2. I'm a bit confused on the mixture that the egg cells are stored in after I'm wondering if the following ratios are correct and if I'm missing anything 100 µl - M2 Solution 125ng - DNA plasmids (ie: pGeneGrip, pcDNA3) 1x - Egg cell 1 µl - Liposomes (Fugene, BoehringerManheim) ???? - mineral oil
  6. Hello everyone I'm currently a student so apologies in advance if I'm incorrect or describe what I'm trying to do poorly, but essentially I'm kind of confused on how extracted mice embryo are fertilized and genetically modified during in vitro fertilization. To be a little more specific consider the following situation and again if I'm wrong about anything below please correct me: 1. A pre-pregant female mouse is euthanized and then dissected to retrieve it's embryo. 2. A mammalian plasmid vector such as pcDNA3 with a cationic liposome coating is added to the embryo so the embryonic cell can uptake the plasmid DNA. One of the things I'm confused about is how the embryo is stored and how the plasmid vector is introduced to that solution. 3. The embryo is fertilized. Not sure how this occurs. 4. Embryo is implanted into a female mouse using an nset device. Any guidance would be greatly appreciated, thank you in advance for your time.
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