Bluenoise

Yeah I get cramps like that if I don't work out. But only if I've been working out regularly. It's weird. Right now i've been going to the gym really hard every second day or so. I know if I take more than one/two days off from the gym I'll get cramps. It's kinda a good thing. Keeps me going lol. I'd suggest trying working out again, but don't hurt yourself. Where do you get your cramps? Mine are almost always in the calves, once or twice I got them in the archs of my feet.

Let me guess your friend is like 13? Or maybe he's just stopped getting smarter at that age. I know I'm 25 and I swear I'm still getting smarter.

Hey great thanks that is what I'm looking for. I've found tonnes of papers dealing with sequencing from beads, however unfortunatley there are some minor technical differences that come to mind between the two. *Edit* Hmmm actually it was not exactley what I was looking for. But it might serve as a starting point. The once instance that this is not carried out in emulsion uses a very small dilution of beads, so settling of the beads insn't a problem...

Okay the last thing you have to do to check this is to observe the sausage while cooking. If the shrinkage occurs prior to heathing stoping then you know this explaination is wrong. If it occurs immediatley or shortly after stopping then it is correct.

"Herbal tea" or "red tea" aren't teas. These are comonly used terms to alternatively refer to tisanes. It may seem odd at first but the word tea doesn't include "herbal tea". Though many commonly missuse it in this fasion. Though personally I rather like rooibos.

Okay, so anyone have any experience with this? Lets say I have steptavidin coated magnetic beads coupled to 500pb Double stranded DNA via biotin. If I wanted to use this DNA as a template for PCR, I am presented with two ways of going about it. 1) Eluting the non-biotinylated strand of DNA, and using it as a template for PCR. 2) Using the beads with coupled DNA as template directly in the PCR reaction. This is preferable as there will be some DNA coupled to the beads that will not amplify and I can remove this contamination using the magnetic properties of the beads. So my question is which method is preferable. What kind of conditions would I need to elute a 500bp sequence from the beads? I'm guessing deinonized H20 at 70-92 C? If I was to perform PCR directly on the beads would I want to include anything to keep the beads suspended for longer? E.g. Poly ethylene glycol? would that interfere with PCR? I know that high temperatures for PCR (i.e. 94C) will eventually destroy my beads by causing them to irreversibly aggragate, however I really only need two cycles of amplification for my purposes so this shouldn't be a problem. Anyways If anyone has any idea's on this, or knows of any papers that would interest me I'd be really happy to hear of it.

I'm not 100% positive, as this isn't a technique I typically work with. However, It likley has to do with primer bias during the reverse transcription step.

We have a winner!!! I'm kinda embarrased I didn't come up with that. It's seems rather obvious now.

Okay here's the speel on tea. There is only one Tea plant. No depened on how the leaves are harvested you get different types of tea. The tea starts off with high levels of citins, however phenol oxidase that is present in the tea converts these to other compounds, such as caffiene So to make black teas the leaves are left to age for a while. The creates lots of caffience and flavor. To make green teas the leaves are friend or steam shortly after picking to deactivate phenol oxidase and prevent this process from happening. There is even white teas, which is the young buds of the plants that are fried right after picking. And it has a very light taste. All teas that don't fall into this catagorie are herbal teas and are calle tisanes. Personally I think any teas with milk, is absolutley disgusinting. I don't understand how someone can put milk or cream into tea. Coffee I understant, but tea... Ewww... I only drink my tea straight or with some lemon.

Lol, am I the only one noticing a theme in this thread. Let me continue it. Which one of these molecules do you think has the greatest degree of conjugated bonds?

He means that the process of reverse transcription doesn't treat every transcript equally. IE some are over represented and vise-versa when converted into DNA. Use of a gene of constant expression can not help with this problem.

Abestoes.

Sure. It's possible it just depends on what charges you're starting with in the first place.

^^ He's so unspecific that your guess is just as good as anyone elses.

Well it all depends on what he wants to do. I don't see how specializing fits into this. Even if he does do biochemistry in grad school he will have to specialize. Most programs are entirley research based, so you specialize by picking your research project. Which can be everthing from working with nucleic acids to tackling mechanism behind cancer. I only have to take 1 course in my program, and it doesn't even have to be a biochemistry course. If on the otherhand he'd rather go into another field altogether than he has many options such as organic chem, medicine, etc. Ask him what topics interest him, and then he should find labs studying that. And apply to whatever program lets him work on that topic. Once you get into grad school the name of your program isn't anywhere nearly as important as the kind of research you do and the papers you end up publishing. Other wise there is work in industry. However without some grad school this is typically limited. You might have luck finding placement as a lab technician. However, this isn't a position that really has any room for advancement. And finding a placement in other industry can be very very difficult. I have some friends who took a long time to find any work. And even then didn't like it or got fired due to inexperience...

Yeah where's that newsletter??

That's easy to answer. What's the Time constant from your electroporation? 4.8 is ideal. However typically anything from 4.4-5.0 will work somewhat. Ethanol percipitation should remove the little salt that's in the gateway reaction, but if you want to remove even more wash your pellet with 70% Ethanol. If I could make a recomendation I'd say to do the following. 1) Make sure that you're doing the LR reaction properly first. 2) Skip the Ethanol percipitation. Just take 1uL of your reaction and use that to transform. The Gateway reaction is so efficient that you'll likely not need to concentrate the DNA at all. (I know they might say to concentrated it to remove things that interfere with transformation, but I usually find this unecessary)

This is just a guess, but as you don't seem to have an fear of these things, and nothing in particular brought it on... You'll likley have two options: 1) Take medication, which may or may not work. 2) Stop playing these games. And really would you actually take drugs just so you can play a couple video games.... Ohhh I recomend NOT playing America McGees Alice in Wonderland....

Yeah I know it's hard to believe that the american public know about this place and actually let it go on... ...But believe me they know quite a bit I think. Stranger things have happened. I believe many see it as "unfortunate and wrong, but a matter of necessity"

Phi is right decimal system was commanded by God. He gave us 10 fingers so we could count to ten. And that is why we have the decimal system We just expanded on it, with 10 10's to give 100.

I was not asking you "What the evidence is" I was asking where the evidence is. Lets see it. All you present are unsubstantiated claims. What is the case study you speak of? What makes it the slightest bit credible? You claim such a massive huge discovery but yet you seem unable to present even a shred of data supporting it. Why should anyone take you the slightest bit seriously? You think that signing your posts with "PhD" is going to trick anyone into believeing you? There are many "PhD"s here, but do you see any of them flaunting their degrees? Common show something substantial. I dare you.

Yeah you see usually when you come up with some crack-pot idea the onus is on YOU to provide the evidence. I'm pretty astonished that someone with a PhD doesn't understand this. How exactley did you get this PhD? Mail order catalog? I'm waiting.

Yeah I thiank we realise that.... I was responding to the comment about vacume chambers and boiling points. I believe that this freezing to remove water from liquors only works on those with a high sugar content. Correct me if I'm wrong but from all my experiences with freezing various solutions of alcohols and water (and there's a fair bit of it ) they freeze uniformly when not in the presence of sugars. No matter if alcohol content is very high or very low. Anyone know why this effect occurs? I mean why the water in a water alchohol solution can freeze at lower temperatures in the presence of sugar? Or why why the mixture exists as a homogenous solution at room temperature but becomes heterogenous once temperature falls. I can ponder a guess but I'd rather see if anyone knows the answer first.

I think you're going to have to be more specific than that. Can you find a link to a paper? or article at least? (To mean it sounds like you're missinterpreting the recently discovered histone code...)

Yep, I use it occasionally to dry unstable compounds. Freeze in liquid nitrogen then put in a thermus and vacume chamber. Wait a while and you end up with a powder. All water can be removed by sublimation. No liquid phase at all.