I want to infect A.thalaiana with a particular plasmid using Agrobacterium but is tDNA necessary or will the Agrobacterium infect the plant without tDNA? And how do I know where, if the plasmid has been inserted?
204045
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.
155640
Tri reagent is a mono-phase solution containing phenol and guanidine thiocyanate.  For RNA extraction, the pH of phenol should be 4 (which helps keep DNA in the interphase following chloroform addition).  Most manufacturers sell pre-mixed Tri reagent containing phenol, so it is likely that you will not need to add additional phenol to your solution.  Check this first.\n\nFor extraction of RNA from plant cells, your method of homogenization may be important and you may need to remove insoluble material by centrifugation prior to phase separation with chloroform.  Also, this is messy stuff and phenol is very dangerous, so I recommend adding "phase-lock gel" to your tube (made by Eppendorf, purchase via Sigma or VWR) to physically separate the organic (phenol) layer from the aqueous layer containing your RNA.
8864
Please don't give your plant a vitamin made for a human. The plant is energetically connected to you now. Just from sharing in your space. It would be in your best intrest to take very good care of the plant. It will bring you prosperity.
238921
Viruses are not free-living...they depend on the host cell machinery to keep them "alive" and allow them to replicate.  Therefore, the only way to cultivate viruses in the laboratory is to grow them in tissue/cell cultures.  You can then allow the tissue/cell culture to incubate and isolate the viruses at a later time.\n\nI don't know what viruses in particular you're asking about, but if you are trying to grow bacteriophage, you can plate the phage suspension on a lawn of bacteria and allow plaques (areas of clearing where the phage have killed the bacteria) to form and then cultivate the viruses from there.
506136
Thymine pairs with Guanine.\nCytosine pairs with Adenine.\n\nThus, the complementary strand would be GGATCGA.
363051
I use DNase I everyday to perform DNase I footprinting studies. These help me determine the binding properties of certain compounds to DNA.\n\nYou can also use DNase II and micrococcal nuclease though they different cleavage patterns.\n\nDNase I can also be used to remove DNA from RNA samples to avoid genomic DNA contamination of samples for qPCR experiments.\n\nDNase I has a preferred sequence-selectivity and does not cut all bands equally. It especially dislikes runs of poly(A), poly(T) and poly (AT).\n\nIf you want to see some images of the applications then just PM me. I've published several papers using DNAse I footprinting so I'm more than happy to share the results.
255756
We did this in a lab I studied in in college.  You have to grow the cells in a stationary location with light.  Often they will differentiate on their own, but you may heve to use a root promoting auxin to get the plantlets to take root.  Once they do, you transfer them to a suitable growing mix and voila! you have a plant.\n\nIncidentally, we pioneered some work growing Niotiana tabaccum cells in suspension by continually agigtating them.  They will not differentiate if you do this.
579911
GM- genetically modified\n\nthese plants have their genes altered and there are many possible mutations that can occur.  also, not much research has been done on the long term effects GM food has on our body.  Why do you think Zimbabwe and Angola rejected our maize offer? because they felt that they were the guniea pigs.  anyway, if you look at how a gene is inserted into a plant using the recombinant DNA method, it is actually very dangerous and unstable.  Scientists are making use of bacteria' plasmids to act as a truck to insert the gene.  Although viral causing genes are removed, what makes scientists so sure that they wouldn't remain, mutate and form a super bug?\n\nAlso, there are religious ethics violated, e.g. some people believe we shouldn't play God and Muslims, they cannot eat pork.  What if the pork gene was inserted into maize? and they eat the maize??? also, some people might be allergic to peanuts, if the peanut gene were to be inserted into a crop, and starts expressing itself, would the person die?
224437
Wow, that sounds like a whole essay. One example is BT cotton which contains a toxin from the bacteria Bacillus thurengensis (spelling?), this prevents cotton plants from being eaten by boll weavils. Valuable/beneficial because the cotton can be raised without having to spray chemical insecticides on the field which can then spread into water supplies. There are another line of crops called Round-up Ready, which allow you to spray weed killer without killing the plants you're trying to grow. You could probably check out the Monsanto website and find some good info there.\nOh- another good one is Golden Rice. It contains genes to make beta carotine to prevent night blindness in children in developing countries
295950
Hypersensitive response-assisting protein (HRAP) has been\npreviously reported as an amphipathic plant protein isolated\nfrom sweet pepper that intensifies the harpinPss-mediated\nhypersensitive response (HR). The hrap gene has no\nappreciable similarity to any other known sequences, and\nits activity can be rapidly induced by incompatible pathogen\ninfection. To assess the function of the hrap gene in\nplant disease resistance, the CaMV 35S promoter was used\nto express sweet pepper hrap in transgenic tobacco. Compared\nwith wild-type tobacco, transgenic tobacco plants exhibit\nmore sensitivity to harpinPss and show resistance to\nvirulent pathogens (Pseudomonas syringae pv. tabaci and\nErwinia carotovora subsp. carotovora). This disease resistance\nof transgenic tobacco does not originate from a constitutive\nHR, because endogenous level of salicylic acid and\nhsr203J mRNA showed similarities in transgenic and wildtype\ntobacco under noninfected conditions. However, following\na virulent pathogen infection in hrap transgenic\ntobacco, hsr203J was rapidly induced and a micro-HR\nnecrosis was visualized by trypan blue staining in the infiltration\narea. Consequently, we suggest that the disease resistance\nof transgenic plants may result from the induction\nof a HR by a virulent pathogen infection.\nVarious defense responses are induced when
441301
Because we never know exactly all the changes the trasgene is doing in the transgenic organism. If its not very well controlled it can be harmful or dangerous. Imagine you put a gene in a fruit tree that codifies a protein important to avoid insects (and you don't have to use insecticides!!). Very good. But you will sell that fruit and someone (not aware of the transgene) might be allergic to that. That's why we need laws and control over that kind of things!
503849
I always did the dephosphorylation simultaneously with the restriction. I used NEB restriction enzymes and shrimp alkaline phosphatase (SAP) from USB. As long as the total volume of enzymes as less than 10% of the reaction (to avoid too much glycerol in the mix) it worked fine.\n\nYou can heat inactivate XbaI. Also SAP is heat inactivated (though I haven't tried it). Check if CIP can be heat inactivated. If it can then probably you can get away with heat inactivation of the mix before using it for ligation. If however you have residual phosphatase activity then ligation will fail.\n\nIt looks like you have a medium or low copy number plasmid, but anyway since it is so big it sticks very well to the column.\nThe best choice would be to prepare lots of plasmid and afford a single purification step, just before doing the ligation.\nWhy don't you just use bigger culture volumes to get more plasmid? You could either do a midi or maxi prep, or use >1 mini-prep columns. \n\nI've been working with very low copy number plasmids so I know the problem. If you are cloning a PCR product I'd suggest that you first clone it in a simple high copy number vector like pBluescript, sequence it and then subclone it to your desired "problematic" vector. Otherwise you might get problems with the sequencing reaction-I've been there. If you are subcloning then OK.\n\nBy the way, the transformation efficiency for very big DNA fragments increases a lot if you add gyrase in the mix. I don't quite remember the citation (could be nucleic acids research) but I bet you can find it on Pubmed.
496967
http://www.google.com/search?hl=en&sa=X&oi=spell&resnum=0&ct=result&cd=1&q=plant+propagation&spell=1\n\nhttp://forums.gardenweb.com/forums/load/propa/msg0212575212518.html?2\nhttp://www.sciencenetlinks.com/Lessons.cfm?DocID=22\nhttp://classroom.all-science-fair-projects.com/forum/showthread.php?t=1161
571684
So you'll need the right Bt strain - Bta or Btk; and not all commercial formulations of Bt contain living cells, so you’ll need some active colonies. I would guess that the nutrient broth that manufacturers use is proprietary. The trouble with entompathogens is that they lose virulence when grown outside of an insect. Manufacturers compensate by developing growth mediums that include insect parts or a blend of certain proteins, antibiotics, nutrients, etc. You can probably find some recipes in research journals but you’ll need to visit a university library or pay for a custom search. You can try Ward's Scientific for culturing supplies http://wardsci.com/  You may also need stirrers or shakers and a place suitable for sterile culture. If your broth gets contaminated with fungi or other bacteria you might just kill all of your bees.\n\nAfter you grow your bacteria, you’ll have to induce them into sporulating. The bacteria themselves do not affect insects; it’s the chemicals used in sporulation. After sporulation, you’re left with a bacterial spore, the husk of an empty bacterial cell and a crystal of toxin (it’s an insect stomach poison with no affect on humans). If done in the proper setting, you can harvest these crystalliferous remains, weigh them to calculate the proper dose and formulate it for easy application. It’s not a simple procedure.
448787
My answer may not be right. I am still a student. If I've gone wrong anywhere, feel free to correct me.\n\nDNA   :  ATA   CGG   TAT   ATG   TTT  TGA   CGC\nDNA   :  TAT   GCC   ATA   TAC   AAA   ACT  GCG\nmRNA:  AUA   CGG  UAU   AUG  UUU   UGA  CGC (assuming the second row of DNA is the template for mRNA synthesis.)\ntRNA   : UAU   GCC  AUA   UAC   AAA   ACU  GCG\n  \nIndividually, this is what each tRNA will bring to the ribosome: Iso Leusine, Arginine, Tyrosine, Methionine or start codon, Phenyl alanine, stop codon, Arginine. \n\nBut you alwayz need a start codon to begin transcribing. Therefore the first three amino acids may not b produced (ie. if transcription hasn't started b4 this sequence). It will start with methionine and stop after phenyl alanine. So the last arginine will probably not be produced either. \n\nAnd the last question.. I dont see any glutamate or valine being coded for here. So I'm assuming I went wrong somwhere.. or I did not understand the question. Sorry, thats the best I can do.
376474
I recommend the following link:\nhttp://www.amazon.com/Phytochemical-Methods-Modern-Techniques-Analysis/dp/0412343304\n\nYou will need to purchase a lab guide or consult with a Life Sciences Dept. for further assistance
225158
I can help you if you need it, I'm a senior biology major. Email my nickname (akidagurl84) at yahoo.com. Hope I can help.