rasing02

I want to prepare 1mg/ml stock of γ-Globulins from bovine blood as a standard for Bradford's assay, I dont know if I should dissolve the powder in distill water or some buffer? Please guide me with the same. I also want to prepare 25U/ml Dnase I and add it in my lysis buffer to lyse proteins. I have a powder form of Dnase which mentions it is 20000 U and wt is 4.12g, I am confused about how to prepare a stock of 25U/ml and I should dissolve the enzyme in what type of solution? water or some buffer or glycerol???

If you can only see the marker i.e your 1st and last lane and not the digest and PCR product that means that nothing is wrong with the gel, something went wrong with your digest and PCR, seems like you dont have DNA in those tubes?? Looks like you did not add dna in the digest or PCR? sometimes that happens...when you are doing a reaction make sure you write it on a piece of paper and tick of every little component you add in ths tube...I hope I helped you. if you are sure you have added DNA then i dont know what the problem might be quote name='Gamewizard' timestamp='1291484085' post='576000'] Hi all, I really need help on this. I recently did a experiment where i ran a gel electrophoresis, and then got a printed copy of the gel image, I had 7 lanes/samples but unfortunately it did not come out all well due to problems with the gel. lane 1 had 1kbp ladder lane 2 had undigested plasmid lane 3 had Ndel digested plasmid lane 4 had hindIII digested plasmid lane 5 had Ndel and hindIII digested plasmid lane 6 had PCR product lane 7 had 100bp ladder So i can only see the first lane i think and last lane/band patterns, but not all of them. How would I prepare standard curves and use them to calculate the sizes of uncut plasmid, single digest, double digest, and PCR product? please help anyone

Why is the homologous recombination event rare in plants as compared to animals or bacteria?

Do you mean Ecoli Plasmid? Various kinds of plasmid can be transformed into cell via chemical transformation or electroporation and similar process, I hope I am answering the question you addressed? Is this what you asked? if not then sorry..i tired

Hi, I am aware of basic synthesis of lipids, I would like some one to help me with the following answer: What is the difference between prokaryotic versus eukaryotic fatty acid synthesis/ lipid synthesis? You could help me with some key differences or direct me to a good paper which compares and contrasts both. Thanks !

I want to prepare chemically competent cells for BL21DE3 ( PLySs) cell line, can someone please email me or upload the protocol for the same, my Id is rasing02@gmail.com, I also wanted to know the protocol for preparing competent cells varies for different cell lines of Ecoli? or remains the same, meaning will it be different for Dh5alpha versus BL21DE3 ( PLySs) , I would really appreciate a quick response.

thanks a ton

Thanks for your response. Why are plants tolerant towards this occurence, while it proves lethal in animals.

thanks a lot, now i understand this concept, so through testing what its needs in minimal medium to grow, i can find what kind of biochemical compound it lacks, right? I really appreciate your help. Thanks !

polyploidy is more common in plants as compared to animals, why?

Law of Independent assortment can be related to anaphase 1 or metaphase1 and how? Law of segregation can be related to anapase 2 or metaphase 2 and how?

I meant let say the unknown gene function is concerning fatty acid biosynthesis then overexpression of this in plant system would not show a phenotypic change? Am I thinking right ??? if not then please correct me, I am finding this difficult to understand. If the gene is invovled in biochemical pathway which leads to production of any of the intermediate etc, then how we will see it via phenotypic change??? Would really appreciate your help.

Thank you, I would look it up and ask any further question if required, this is mainly going back it how it all started, when technology was not there how people found the function of unknown genes Merged post follows: Consecutive posts mergedReverse genetics can help in determination of a specific phenotypic change, how will it help in determining biochemical/metabolic change which is not phenotypic??? Merged post follows: Consecutive posts mergedThanks for thr response.Reverse genetics can help in determination of a specific phenotypic change, how will it help in determining biochemical/metabolic change which is not phenotypic???

1)If a particular plant seed has anticancer properties then how will you find which phytochemical is are responsible for it? 2) How can we test various factors affecting the production of phytochemicals and how can we indentify all the genetic factors responsible for synthesis of a phytochemical 3) how can pytochemicals be produced in a recombinant system?

If you have a gene sequence which is known to be expressed but does not have a biochemical or physiological function, then how can one determine the function and prove it, lets say in a plant system? Also consider the fact that you have to work from scratch no database (gene bank, blast,etc) is avaliable???