How necessay is tDNA if I want to infect A.thaliana with a plasmid?
452878
From my understanding tDNA is important, at very least for tagging but I believe it is also important for the transfer of the intesert into the A. thaliana genome.\n\nYou can still detect the insertion of the transgene because you would usually insert an antibiotic resistance gene with the gene of interest. You can then use this to select for plants that have the gene inserted.\n\nThen, to find out where in the genome the transgene has inserted, you need to map it. There are many techniques for doing this, but if you know the sequence of your gene of interest or the antibiotic resistance gene, you should be able to use fragments of the genes as probes on gene libraries etc etc etc, there are probably also fancy ways you could use genomic to achieve this too.
226604
http://www.washingtonpost.com/ac2/wp-dyn?pagename=article&node=health&contentId=A45289-2000Apr9
349513
http://www.altavista.com/web/results?itag=ody&q=procedures+requiring+sterile+technique&kgs=1&kls=0
729453
K.C.'s answer works if you have a selective media that will work. Also (or in addition to that) you may want to plate out colonies and then PCR out a unique sequence or two.\n\nThis wouldn't be definitive but if you design your primers right it may work, but also may be very time consuming.\n\nAlso you may want to try restriction digests to remap your gene. This would work well, if you have a good map.\n\nGood luck, what a bummer
843667
How is Recombinant DNA made? \nThere are three different methods by which Recombinant DNA is made. They are \nTransformation, Phage Introduction, and Non-Bacterial Transformation. Each \nare described separately below. \n\nTransformation\nThe first step in transformation is to select a piece of DNA to be inserted \ninto a vector. The second step is to cut that piece of DNA with a restriction \nenzyme and then ligate the DNA insert into the vector with DNA Ligase. The insert contains a selectable \nmarker which allows for identification of recombinant molecules. An antibiotic \nmarker is often used so a host cell without a vector dies when exposed to a certain \nantibiotic, and the host with the vector will live because it is resistant. \n\nThe vector is inserted into a host cell, in a process called transformation. One \nexample of a possible host cell is E. Coli. The host cells must be specially \nprepared to take up the foreign DNA. \n\nSelectable markers can be for antibiotic resistance, color changes, or any other \ncharacteristic which can distinguish transformed hosts from untransformed hosts. \nDifferent vectors have different properties to make them suitable to different \napplications. Some properties can include symmetrical cloning sites, size, and \nhigh copy number. \n\nNon-Bacterial Transformation\nThis is a process very similar to Transformation, which was described above. The \nonly difference between the two is non-bacterial does not use bacteria such as E. Coli \nfor the host. \n\nIn microinjection, the DNA is injected directly into the nucleus of the cell being \ntransformed. In biolistics, the host cells are bombarded with high velocity \nmicroprojectiles, such as particles of gold or tungsten that have been coated \nwith DNA. \n\nPhage Introduction\nPhage introduction is the process of transfection, which is equivalent to transformation, \nexcept a phage is used instead of bacteria. In vitro packagings of a vector is used. \nThis uses lambda or MI3 phages to produce phage plaques which contain recombinants. \nThe recombinants that are created can be identified by differences in the \nrecombinants and non-recombinants using various selection methods.
1135140
Not a good idea.  If you are using water from the faucet and for that matter even sterile water there is chance that you will get an infection.  Imagine injecting 20 cc of water into you biceps and the water has germs.  The infection could be quite nasty and quite painful.
460908
human don't have plasmid. but if you're asking how to test a bacterium for human growth hormone (hgh) recombination into its plasmid, you can perform is PCR test for the HGH mRNA. that should be the most direct proof.\nor you can do a DNA sequencing, but that takes longer.
181048
deoxyribo nucleic acid
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I gave a lecture on this recently and have had first hand experience with this technology in a lab, so let me give you a brief and easy summary to take with you about this topic about Recombinant DNA Technology:\n\nIn order to clone you need 2 things: Bacterial DNA (the circular form called a "plasmid"), and the DNA you want to clone (be it human, mouse, plant, other species etc)\n\nThen you will need to cut the Bacterial DNA with enzymes called "endonucleases" that work like scissors. (So imagine a circle and I take a pair of scissors and cut the circle at some point in it, b/c I want to insert the foreign DNA (non native DNA) where I had cut.) I will have to do the same with the foreign DNA and have endonucleases cut OUT the particular piece i want cloned. \n\nThen we can put the two pieces together, the cut circular plasmid DNA from the bacteria, and the CUT out piece of foreign DNA. \n\nWhen we put the two pieces together it is called "ligation", and it is done as a reaction in a tube with enzymes inside that "glue" the "sticky ends" of the bacteria DNA to the foreign DNA. \n\nAfter the reaction normally done in a 16 deg. Celsius water bath, we now will have "circular" plasmids with our Foreign DNA inserted into the bacterial DNA (the circle DNA)\n\nThen we would re-insert this DNA back into the bacterial host (that the circle DNA first came out of) in a process called "transformation", this is where the broth from the water bath we just discussed is grown in a growth medium at 37 deg. celsius so that the bacteria can grow and produce MORE of those circle plasmids with our foreign DNA inserted into it. \n\nThen we would "plate" these onto agar plates, and let them grow in incubators at 37 deg. Celsius over night until we get small colonies of bacteria on the plate. \n\nEach "mound" of bacteria (small like white dots on the plate) contains millions of copies of our gene (inserted in the circular plasmid DNA of the bacteria). \n\nNow we can take these small colonies, and harvest them further, by putting them in large Erlenmeyer flasks to be grown in the same "growth broth" over night on a shaker for 37 deg. celsius. \n\nAfter of which we can harvest, and purify that culture so that only the circular plasmids remain (this is a long and convuluted process so we won't go into it in detail here), and from there you can store those in a 4 deg. Celsius fridge and keep it in stock for further research purposes. \n\nHope that helps!
756837
It is a process by which a plasmid is used to import recombinant DNA into a host cell for cloning.  Differnet companies have developed their own exclusive means to accomplish this task for the reasons listed below.\n\nMany diseases are caused by gene alterations. Our understanding of genetic diseases was greatly increased by information gained from DNA cloning. In DNA cloning, a DNA fragment that contains a gene of interest is inserted into a cloning vector or plasmid.\n\nThe plasmid carrying genes for antibiotic resistance, and a DNA strand, which contains the gene of interest, are both cut with the same restriction endonuclease. The plasmid is opened up and the gene is freed from its parent DNA strand. They have complementary "sticky ends." The opened plasmid and the freed gene are mixed with DNA ligase, which reforms the two pieces as recombinant DNA.\n\nPlasmids + copies of the DNA fragment produce quantities of recombinant DNA.\n\nThis recombinant DNA stew is allowed to transform a bacterial culture, which is then exposed to antibiotics. All the cells except those which have been encoded by the plasmid DNA recombinant are killed, leaving a cell culture containing the desired recombinant DNA.\n\nDNA cloning allows a copy of any specific part of a DNA (or RNA) sequence to be selected among many others and produced in an unlimited amount. This technique is the first stage of most of the genetic engineering experiments: production of DNA libraries, PCR, DNA sequencing, et al.
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It will depend on a number of things.\n\nFirst:  you can plate them on media without antibiotics and they still might conjugate, there is nothing magical about antibiotics that tells the bacteria time to make a sex pilus and have at it.\n\nSecond:  what are the mechanisms of resistance.  If the streptomycin resistance is a mutation to a chromosomal ribosome subunit-encoding gene (normally how that works), then it wont initiate conjugation, unless it happens to also have a Hfr plasmid integrated into the chromosome at the same time (which could then initiate conjugation of the entire chromosome).\nIf ampicilin resistance is due to a plasmid-borne beta lactamase gene, and that plasmid happens to be a F plasmid, or other conjugatable system, then it can potentially initiate conjugation.\n\nIf both resistances are contained on conjugatable plasmids of the same origin then they will most likely be incompatable with each other, and no conjugation will take place.\n\nThird:  Are we talking about the same species of bacteria or different species of bacteria?  It shouldn't be assumed that two different species will readily conjugate.\n\nFourth:  (This is more a theoretical concern than one that I know to be truly an issue)\n\nHow does the antibiotic work?  Lets say in your example, you have 1 strep resistant strain of E. coli and 1 amp resistant strain of E. coli.  If you plated in the presence of streptomycin, the amp resistant strain wouldn't be able to synthesize new proteins, and if there weren't cells already producing conjugation proteins prior to plating, then there won't be any conjugation either, as the streptmycin inhibits protein synthesis.\n\n1, 2 and 3 are concerns I know to be valid, 4 maybe is maybe isn't but occured to me as I was typign the rest of it.
826034
One tRNA is needed for each tri-nucleotide in the RNA that is being transcribed, so in theory 64 would be needed to ensure any RNA could be translated. However there is some bias in codon usage (some tri-nucleotides are not found in certain species) so in practice you would need slightly less than this. Of course if the protein being translated did not contain all 20 amino acids less would also be required then
569271
Plasmids are little loops of DNA. They can act as vectors ("carriers") in genetic engineering. The donor DNA is inserted into the plasmid/ vector, which can then be inserted into a bacterial cell, typically a non- pathogenic species. This will transform the cell. Large numbers of these bacteria are then produced in conditions which favour synthesis and secretion of the desired gene product. \n---> Important for the artificial production of hormones etc.
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You have to identify two sequences that are >6kb apart from one another that flank the sequence you are attempting to amplify. Better yet, you can get you a well understood cloning vector (plasmid) that already has primers develped for it that flank the polyrestriction site region (ie. puc19 [see link below], find the needed restriction enzyme sites that flank your desired 6kb sequence and clone the DNA into the vector. Be sure to check your sequence for any internal restriction sites. This may require you to do a partial digestion and recovery.
71310
http://www.nlm.nih.gov/medlineplus/ency/article/000704.htm
210434
urban legend!!!! Replacing a tesicle with an onion would probably cause a pretty bad infection.
1002331
isolate the pathogen...