Biochemistry and Molecular Biology

I am running a restriction digest with the Pyp04 plasmid (has 1 restriction site for BglII) and BglII. Using 2ng of plasmid, I was told to use 5 uL of BglII to digest. After 2 hours of digestion I ran a gel to test how well my sample was digested, but did not include an undigested sample (I am under the assumption you wont just see one band if it was under digested). The bands were a little on the light side, so I am doing an etbr soak to brighten the gel up. In the meantime (1.5 hours after I started the gel), I was told to add another uL of enzyme to the digestion for another hour. So my question is, would it really be necessary to do so? A typical digestion of th…

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

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???

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?

Im doing a coursework, im at GCSE level. This coursework is late:-( and this doesnt really need to be included in it, but I was thinking about it. The investigation is simple: How does light intensity affect the rate of photosynthesis? We used the classic method, of pondweed in a beaker of water, with a lamp shining at it, from varied distances for a minute at each. To make sure no other factors affected the rate, we took soem measures. One was to add a spatula full of sodium hydrogen carbonate, to prevent carbon dioxide being a limiting factor to the rate of photosynthesis. what i wanted to include in my coursework was exactly how and what reacts, releasing the ca…

Hi everyone, I'm hoping someone here has used HRM to do SNPs. After reading some of the literature, I am a little confused about a few things, and if anyone can answer any of these I would very much appreciate it! - can HRM only be used for one SNP at a time? Is there any way to multiplex? - do you need special primers, or can you use any of the ordinary primers you can order online? - is this really better than real time PCR to do SNPs? Speaking of real time PCR, can you do more than one SNP at a time that way either? Thanks for any insight anyone has!

As I recall back from my high school health and biology classes, the human body produces insulin in order to regulate glucose. However I read recently that insulin stops the use of fat as an energy source. I was amazed by this information. I used to consume copious amounts of sugary (high fructose corn syrup actually) drinks. Since reading this I have cut back drastically. I want to know more about this. Are there any data showing how long it takes (in non-diabetic individuals) for insulin to lower back to the "fat burning level"? For example, if I consume a 12oz can of Coca Cola, how long will it take for the anti-fat burning effects to wear off?

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?

I'm reading about pyoverdin, and I've decided it seems very similar to GFP. Why were researchers in the past so focused on GFP as a marker in cells rather than pyoverdin? Was it that GFP better enabled itself to be embedded in proteins rather than be used an a gene insert?

Hi all. This is my first post here, so: 1. I've read the sticky and think this belongs here and not in Speculation. 2. I am a scientist of no sort, so my question may be face-palmingly stupid. That said, what are the limits of bacterial synthesis? I understand that bacteria have been bred that synthesise biofuels and pharmaceuticals, but if the methods available were flawless, could they be engineered to create things like foods, explosives and varieties of plastic? If so, what would they need to feed on? I doubt they can make everything from agar and glucose.

Hello everyone, I'm struggling a little bit to understand the structure of haemoglobin. I have few questions to ask. 1.Ok so the top bit must be the proximal histidine and in haemoglobin rather than water bottom bit is attached to distal histidine when oxygen is not present right? Now my question is why iron atom able to make 6 bonds. For example if it is oxygen, I know it has 6 electrons in final outershell so it need 2 electrons meaning 2 bonds. (I know energy levels are bit more complicated but this is my level at the moment). Using the same prinicple iron has 26 electrons, that mean it has 16 electrons in its furthest shell so I don't understand where 6 …

I don't really have biochemistry experience in this so I was hoping someone could give me suggestions. I have isolated an enzyme from whole animal tissue, but it lacks all activity. I ran a couple of native gels and it looks like it is denatured (bands appear the same as those boiled in sds buffer prior to loading). I have a paper where they refold the protein using urea, but they are refolding inclusions bodies from e. coli overexpression (long story why we use tissue...). First question, does refolding even work for proteins isolated from whole tissue? Almost all of the papers I have found are always working with proteins generated in ecoli. If it's still doable, I'll a…

Hello Scientists: I was fishing on internet for companies who sell bacteria cultures (5-6 different strains) for our experiments, butr no luck! Our experiments include detection of spore forming cultures based on biomarker called dipicolinic acid (DPA) using mass spectrometry. We need 3 different strains of Bacillus and Clustridium species, which from endospores and contain DPA, and as well as 3 different strains of bacteria which do not form spores. If anybody has purchased such cultures from any company, would you PLEASE provide theit contact information, or at least the name of thet company? thank you very much in advance! frank

Hi guys, I have a big question in my mind, given that I am not an expert biochemist. How difficult is the isolation of big RNAs? I'm thinking on lengths between 2 kb to 20 kb, or even larger. Besides, these RNAs have very low abundances. Could we faithfully perform in vivo analysis of their function, like immunoprecipitations? Any help would be great. Thanks!! -Sakti

What is the origin of a spermatogonium cell?

Hi guys, I'm wondering if there's a way I can get the full sequence in ACTG format for the human pro-insulin gene INS1? It's on chromosome 11. With introns would be good but I'm not familiar with the online databases

I want to sequence a 47 nt RNA on a 40cm long gel plate. What is the percentage of the acrylamide should I use to resolve all the 47 bands on the single gel at the same time? And what is the difference between Acrylamide:bisacrylamide ratio 19:1 and 29:1? which ratio is appropriate for me? I tried 12% 19:1 Acrylamide : bisacrylamide gel. the bands between 35 to 47 were not separate. Thanks a lot

Beta-D-glucose methyl-alpha-D-glucoside alpha-D-fructose sucrose maltose and also what is the number of the carbon atom that is an aldehyde or ketone group in open chain form?

Web: http://jcmb.halic.edu.tr E-mail: jcmb@halic.edu.tr • Journal of Cell and Molecular Biology is an international journal which covers original research works in the field of cell biology, molecular biology, genetics, microbiology, neurobiology, bioinformatics and related topics. The Journal aims to encourage publications with an interdisciplinary approach. • This journal publishes research articles, review articles, short communications, book/software reviews, case reports and letters to the editor. • Contributions are published in English and in Turkish. • Independent peer-review system guarantees that all studies appearing in Journal of Cell …

Hello all. I am having problems with the X-gal assay of my yeast two hybrid. The assay has been done before by a former lab technician, and despite following her instructions, I am unable to repeat her results (which she was able to repeat). I am re-doing the "confirmation" part of the yeast two hybrid protocol, in which I transform in my prey plasmid (from pACT2 library) into naive strain Y190. Next, I transform in one of several bait plasmids (vector pBG4D); these include vector only, and several constructs of our protein of interest (different domains, full length). I am successfully able to transform in my constructs; they all grow on selective media (in my…

Hello, my name is Justin Collier and I am a high school Biology student. I am currently working on a report on the potential use of DNA as a computational machine. For the report I need to have interviewed at least one person who works in the field. I have familiarized myself with the research of Ehud Shapiro and have emailed him asking for an email interview. Does anybody know of other leading figures or researchers involved in this research? Thanks, -JNCR

I'm trying to write my lab report up and I'm getting really confused about a certain calculation! I made a 1% agarose gel, and Basically, I added 5 μL of ethidium bromide to a solution (solution= 0.5 g agarose and 50 μL TAE) and the bottle said the ethidium bromide had a concentration of 5 μg/μL. so what concentration of ethidium bromide am I adding to my solution? Help would be much appreciated!

1 to 4 are arabidopsis leaf ,5 to 8 are arabidopsis root why there are so many bands in leaf tissue,and what is the band in front of 28s rRNA

Is it possible to create artificial cell markers? I was thinking that such a thing would be great for artificial hearts and the like. Iirc, recipients of artificial hearts have to take immunosuppressants. Would an artificial heart that the body thinks is its own relieve that need?

Hello people I found myself dealing with this experiment... In my school, we've set 7 tubes with the following quantities of sucrose at pH 5.0 (umol): T1 = 0 T2 = 5 T3 = 10 T4 = 25 T5 = 37.5 T6 = 45 T7 = 50 So after climatization step at 37ºC, we added same quantities of invertase to each tube and took it to water bath at 37ºC for 15 minutes. Then we added DNS and took the tubes to boiling water bath so the enzyme could denature. After their cooling off, we took the tubes to spectofotometer so that T1 was the blank and the absorbance data was taken from the others. For the ones who don't know, point is that this enzyme forms reductor sugars (glu…