Meagan's Friends' Journals
Thursday, May 15th, 2008
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sunnybananas |
5:07a Cast Away
(Posted "nightly", around 5am, using LoudTwitter) |
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_scientists_ [ gnommi ] |
8:56a Protein Quantification Hassles I've just started doing Western Blots to verify some immunohistochemistry results but I'm having a few problems. On my first attempts I used NP40 or RIPA basic plus protease inhibitor cocktail as my lysis buffer, but got rather low protein yields. This time I used SDS/Tris-HCl with Roche Complete protease inhibitor cocktail as lysis buffer (made up as per the WB protocol on the antibody data sheet), which gave a much more viscous lysate. However, when I went to quantify my protein yield with my Fluka Rapid kit (basically a Bradford assay with Coomassie, and BSA standard), every time I got a horizontal line instead of the usual curve with my standard dilution series. Even by eye I could tell that the optical density of all of my standards was the same (not halving between dilutions as it should). Could my kit be out of date? (There is no expiry date on it, and it's been stored at 4C). Or do you think the lysis buffer is incompatible with the kit? **annoyed** |
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uog [ grrl_next_door ] |
12:01a DE anyone know if ALL DE courses are "fail the final exam, fail the course"?? Regardless of how well you did on the other stuff? When I took another DE Psych (death and dying) I believe this was the rule and don't know if it applies to the DE Abnormal psych or not... In both courses the final was worth 45% so I dunno. If you could let me know that would be great! Thanks |
Wednesday, May 14th, 2008
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_scientists_ [ mdunnbass ] |
8:37p T4 PNK troubleshooting Hey everyone - So, does anyone know if low pH inhibits T4 polynucleotide kinase? I know that ammonium and phosphate ions can inhibit it, and in the absence of ATP the enzyme has both 5' and 3' phosphatase activity, but no word on pH from what I can find. (I'd call NEB tech support, but it's after 2000hrs, and they are only open until 1700hrs) I have tried labeling the same set of oligos several times now with no success. I know that this particular aliquot of oligos has been successfully labeled in the past, so it's not an issue of having added the wrong DNAs. (I am troubleshooting for DNA degradation though) I know that the buffer that NEB supplies is pH 7.6, and the enzyme itself is stored in pH 7.4, but when I am setting up my labeling reaction, our Millipore water is pH ~4.5, and I am thinking that this might be the reason my labeling hasn't been working in a while. I've ruled out the enzyme and the buffer, so now I'm checking my oligos and the water. I doubt the purification columns could be the problem, since they are fairly fresh, but I'll check on those as well. I do know that when I label and then column purify the labeling reaction, the columns are screaming hot on the geiger counter, and the CPMs of my 'labeled' oligos on the scintillation counter are essentially 0. So, the 32P isn't eluting. Either my DNAs are degraded, or the DNAs aren't kinased, or the DNAs are stuck to the columns. I could see pH potentially affecting all 3 of those scenarios. Any thoughts or suggestions would be appreciated. Thanks. Matt |
Thursday, May 15th, 2008
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fd_midori |
12:30a Paris photos. Street Art, Shibari homo art, trains stations, etc. Just a few odds and end photos... View from Eurostar. I think on the UK side, but there's a section where both sides look really similar. Yellow field is mustard flowers...  See ![[info]](http://p-stat.livejournal.com/img/userinfo.gif){kind=small} kumimonster working hard on her site updates. Gare Du Nord, Paris  ( Read more... ) |
Wednesday, May 14th, 2008
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fd_midori |
11:55p London notes - the joy of the workday Similar to when I'm in NYC, I don't feel like a visitor in London. Not that I know these cities like the back of the hand... but because here my days are spent as a person going about their work day. My hide-away pals treat me like a room mate. We see each other in our robes in the morning, mumble greetings and offer the other coffee. I dress in my teacher's duds, grab my briefcase and join the stream of people on the tube with my Oyster card and head into town. I have meetings, grab a cuppa, text message on my UK phone with friends and text for work coordinations, then I teach the classes and maybe grab a brew with friends after work. Maybe I get to carve out a bit of time for a museum or a garden. The weekends are coordinated with friends on what to do. I stress on deadlines. I read my favorite papers, trade rags, Time Out or listen to the iPhone on the tube. I walk out of the tube, nearly the last train, to the quiet suburban streets and head home to companions for some tea and telly. We complain to one another on the cost of living. It's a strange little comfort here, and New York, for me to feel so "settled in". I just slide into it. (The down side is that for all the years of coming to London, I've not really seen the rest of the UK!) This trip, even the experimental trip to the little fetish party in Paris on the Eurostar felt like a local's weekend jaunt. That kumimonster, my long time pal originally of SF, occupies a flat here part time and we text each other and hang out from time to time, only adds to the nomad's fantasy of a settled life.Some highlights this trip, so far, that make me feel like I just slide into life here. Jet lag was brutal with the uneven sleep schedule the first few days in London / Paris. Really great classes at Coco De Mer! All sold out. More students. More journalists covering them! Productive meeting with a friend, who is similtaneously located in London and LA and I just saw in LA a few weeks back, to brian storm the Big Picture... meeting with a corporation considering hiring me for a special presentation... meeting, just talk, with an agent... Getting thanks from spouses and partners of the students the day after! Hanging out and working on my laptop with great coffee and free WiFi at Coffe Cake and Kink between time. Belgo (but haven't been able to stop by Neal's Yard Creamery or scones at the Covent Garden Hotel yet this trip) Might get to go sailing this weekend on friends' boat to Isle of Wight. (I need to get some equivalent of deck approved shoes.) |
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fd_midori |
11:51p Tigger's results. Monday night K took Tigger to the emergency pet hospital. They waited a couple of hours... then all sorts of tests. Then more waiting. Nothing wrong with her mouth or teeth. Kidney count's slightly elevated but not out of the ordinary, apparantly. She had a fever and was dehydrated so they gave her sub cutaneous hydration. They can't figure out what's wrong wither. After the hydration she's feeling a bit better. Purrs to K's pets. Growls at King David as is customary. She's not eating much kibbles but has been given the OK for boiled chicken and tuna with tuna water. She's eating the latter. Keeping fingers crossed. |
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uog [ twilight_tipsy ] |
5:27p I don't know if this is allowed... I'm moving onto residence in september and I don't really know anyone else who's going to UOG.... i was hoping maybe I'd find someone who is LGBQT tolerant, open-minded and outgoing to double with me ? |
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tousledelegance |
1:37p On my own, but not single I have come to the conclusion that I have lived alone for so long that I cannot live with someone else, no matter how much I care about them. I am too set in my ways. It comes from being spending so much time by myself all my life. Growing up, no kids lived in my neighborhood and my siblings were a lot older and out of the house the majority of my life. I entertained Me and did what I wanted, when I wanted, how I wanted (within reason – my parents were there for me and gave me guidance, don’t go assuming I was neglected or mistreated). I don’t resent having that kind of childhood, because I think it helped foster my creativity, but I will be the first to admit it also had some negative effects on my social skills and gave me some unrealistic expectations about the world. Basically, I don’t really like to be around people. I don’t know what to talk about with them. I’m not a good listener, I don’t know how to ask the right questions and I don’t like to talk about myself. I either wind up angry, paranoid, insecure or frustrated in conversations. Generally, the things I am interested in, most people are not into… likewise, I don’t particularly care for what the things they enjoy. Stalemate. Besides, my main interactions take place at work, where topics like sex, religion and politics are a no-no, or in bars where you usually get what I refer to as ‘inane bar chatter.’ (It seems as though no one talks about anything of substance anymore, but that’s another post in the making.) That isn't so say there haven't been exceptions, nor to imply I don't love my significant other and value the time we spend together. However, in cohabitation, the little things annoy me. (Probably more that they should, but again, consider my background. ) For example... When I put something somewhere, it is for a reason. I want it to be there when I come back to it. I don't want to have to look for it every single time I need it. It wastes time and it makes me neurotic. (And, yes, I am a girl, so there will be girly products in the shower/tub.) I'm sorry if others don't like that, but it akin to OCD for me- a habit I can't stop. I'm tidy- meaning everything has a place, but not neat, however, I'm not a disgusting slob. Sure, I tend to clean when it's critical (read: in the way or cluttered looking) or when company is coming. I respect others space by not leaving gross, filthy, or unsanitary items around to attract bugs, stink or spread germs, BUT I use disposable cookware/dishes/utensils for convenience, I don't want to wash something immediately after using it and then put it away as soon as dry, I might not always make the bed, I might leave the laundry in the dryer or basket for a while, I keep my receipts because that is how I balance my checkbook, I use catalogues and coupons, I sometimes set stuff off to the side so I can go through it when I'm not so busy or exhausted, I use the internet a lot because it is relaxing and it is a means of communication with which I am comfortable. All these 'routines' are my way of de-stressing; it is often the only thing in my life that I can control, just enough to keep my sanity. I'm not well mentally and it sometimes makes me ill physically. I'm sorry I can't afford the treatment I need, but I don't like to be nagged- that only makes it worse. There are days that are good days; those are when get tasks done quickly and without fuss. Other days, I am struggling just to stay alive. Literally. I'm tired. Tired of apologizing. Tired of being made to feel guilty for everything. Tired of feeling this way. Tired of being tired. I was happier living alone where no one told me what I could or could not do, when I could or couldn't do it and how it was to be done. I feel like what makes me Me is being held down and gradually stamped out. I've made multiple attempts to explain how my feelings and to find a compromise. Just when I think one has been reached, the agreement is violated and someone else's breach is always justified by blaming me. It is not a matter of 'I won't live with someone' anymore. I CAN'T. My solution? Nothing as drastic as to break up, but to live on my own again. Nearby. I can stay over once in a while; go home when I need some space. I hope that is fair to everyone. current mood:  indescribable
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_scientists_ [ luciddork ] |
10:21a Journal Access I find myself soon to be graduated and gleefully headed off into the private sector. I realized the other day, though, that I will soon be losing access to all the journals my University has subscriptions to. Is it common for companies to have accounts with journals? Do they buy them on an as needed basis for related research only? What are the common practices for this? In short, to those of you outside of a university setting: How do you read your journals? |
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_scientists_ [ sdjma ] |
11:38a Article Request Thanks!!! |
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_scientists_ [ chemrocks ] |
12:24p TISSUE CULTURE HELP Through an unfortunate accident, I accidentally plated ovarian cancer cell lines to petri dishes. They are adherent cells. Some of them have adhered to the bottom of the plate. I would like to know if the floating cells would still be alive after 2 days if I were to replate it to tissue culture plates now and whether there would be any problems, ie. altered cell properties from being plated on petri dishes initially? Thanks! |
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sunnybananas |
5:06a Storm Clouds
(Posted "nightly", around 5am, using LoudTwitter) |
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dalekboy |
11:28a Back from outer space? No, not really... Because everyone's doing it and I'm a mindless sheep... ( Which Shakespeare Play am I? ) current mood:  tired
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Tuesday, May 13th, 2008
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mycology [ freeofthought ] |
1:21p Shiny gold mushrooms Unfortunatley I didnt pick one to look at the spores, but I found these in Indiana growing on wood; and Ive only ever found them in this one particular place.  
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