See, I agree with everything here. But that’s the point of Taq: you shouldn’t be cloning with it, or for that matter anything more than standard check PCRs. If you do, then you are asking for trouble. The first option is to send the stuff back; $400 of anything is still more than an eyeblink (even though I do burn through faster than I should) and the biotech companies usually are understanding about replacing it. If not, though, then run a few controls and see if it still has activity. Label as “Bad Taq” and use it for checking cloning inserts and plasmid-template non-cloning PCR. It will still get used; I know at least 50% of my PCRs are for colony checks and cloning confirmation. I always run a positive control so I would know if the PCR failed, so I would have no hesitation in using Taq which I knew worked but perhaps not with full activity.
I only use Taq for products <2 kb that don’t need to be cloned. I use Pfx for cloned products <5 kb and Herculase for products >5 kb. I have occasionally used homemade Taq/Pfu cocktails for more difficult transcripts but I find glycerol and DMSO go a long way to chewing through long stretches of high GC.
I have some personal experience with enzymes stored at the wrong temps. In June 2001, a giant flood hit our medical center and we lost power in our building for upwards of two weeks. Emergency power came back on during the third day for our building but stayed off in some buildings for the whole duration. This meant that every grad student that I know had dry ice burns on their arms for months afterwards as it became a twice-a-day ritual of hauling up dry ice to stuff freezers filled with irreplacable reagents (mostly ES cell line around here). Every enzyme, every buffer kept even at room temp (our lab was at around 31 deg C for two weeks) was marked with a red X for testing and disposal. Insurance reimbursed us for many of these, but we still kept some around and tested the activity (especially in the more valuable and harder-to-get stuff). I can tell you that around 3/4 of the stuff worked: random prime labeling kits containing heat-labile DNA pol, restriction enzymes, modification enzymes, RNases, DNases, antibiotics, that decade-old Taq, etc. In fact, we still have some around that we still use (we have a shitload of ampicillin and paraformaldehyde at 4 deg C that we are still working our way through).
Neither would I, but apparently my boss doesn’t share our concerns. We have had some rare restriction enzymes in glycerol blocks on the (worst) top shelf of frost-free freezers since 1994. I am usually suspicious about them, but in my time here, 1 in around 10 that I have tested (that bitch AccI) of them has lost activity (even with the 2001 incident). Not that I would be enthusiastic in using them for cloning or anything quantitative, and not that I would advocate doing this, but it has gotten me through more than my share of test cuts and confirmations in my 5.5 years of grad school. Hopefully I can defend and truck my ass out of here by March…
I think people would sympathize about the specific problem. But if he didn’t have any other grievances with her, pitting monstro personally probably wouldn’t go over too well. “Dude, calm down. Everybody makes mistakes. Is she generally a competent worker?..Then don’t dwell on one mistake.”
monstro , I am coming late to this thread because I thought from the title someone was just pitting a coworker and kept skipping over it. I opened it yesterday and have been thinking about some of your statements off and on since then.
You sound so much like me when I was your age. Even though I got all of the jobs I applied for and raises and promotions I always felt like it was all smoke and mirrors and that at any time these people who thought I was wonderful would figure it all out. I, like you, was not depressed and did not have panic attacks, but I suspect I did have what I know now to be just “free floating anxiety”, ie, anxious about everything with nothing to really pin it on, no confidence in my own real abilities. Somehow I got through that stage of growing up. I think a lot of it came with motherhood. Why I say all this now is to urge you not to just let it go like I did. You are smart and eloquent in your observations. All the people who have posted here know that and see the “real you” and we like what we see. Go find a good counselor to talk to so you can enjoy the real you. You owe it to yourself.
Mistakes? Hopefully you will keep making them until you are six feet under. Show me someone who never makes mistakes and I will show you someone who is not actively engaged in life. And when you learn the lesson of your mistake you can let it go.
I want to share with you something I remember from probably the best boss I ever had. She was Director of Nursing and my boss many, many years ago. I went to her with a huge problem, the mistake of the century, would cost my hospital lots of money. I was beyond anxious and she could see my acute distress as soon as I walked into her office and started talking. She interrupted me early on into my explanation:
Boss: “Did anybody die?”
Me: “Oh…no.”
Boss: “Then I bet we can fix this. Now tell me the rest.”
We could all do worse than be a little like her.
Keep us posted on how things go for you. And if you decide to marry Wendall Wagner we all get invites to the wedding. Hey, think twice before passing up a man who doesn’t mind if you don’t cook or clean.
I once worked for a psychological help center. They started a selfhelp group for people who had had a loved one commit suicide, and I was assigned the task of putting an ad for it in the paper, featuring the telephone-number of the center.
I gave the wrong number.
Twenty people, all traumatized by a suicide, called the number of some rightfully impatient and annoyed guy.
Your post reminded me of myself in so many ways! Judging by the replies here I am guessing there are a lot of people who would be considered ‘smart’ by the outside world who find themselves making boneheaded mistakes all the time. I know I do.
My thought on the subject of my own incompetence is that its the byproduct of a creative mind. I’m always thinking of something else, always dreaming a new dream, always seeing reality in just a slightly different shade of orange than everyone else. Of course I make mistakes during every day life… my attention is hardly ever focused there
In most of the working world secretaries to do mundane administrative tasks for technical and scientific workers have gone the way of the Selectric. It’s interesting…I’ve just read the 1950s classic The Man In The Grey Flannel Suit, by Sloane Wilson, and it’s interesting how his character, at a job that paid $7000 a year–which was like perhaps $70000 today, had his own secretary.
My group has one executive assistant for a group of twenty-odd people; she is basically the secretary for our boss, keeping his appointment calendar and doing other secretary-type. I doubt if she takes dictation or types, since people who cannot type are rare as hen’s teeth these days. Certainly nobody else in the group would ask her to do something like that either; if you’re expecting a package or a visitor you handle it yourself.
So I don’t think it at all surprising that the OP had to sign for a package. She just happened to be there when the delivery came.
I really don’t know why you keep saying this. If you are missing the bottom portion of your face, then kissing might be difficult, yes. Did someone tell you that you aren’t a good kisser? If it just that you haven’t kissed someone, believe me, those things take care of themselves. Besides, keep practicing! Saying that you can’t kiss is like saying that you can’t shake hands.
My husband and I met over our computers. I made it very clear to him that I was not at all interested in cooking or cleaning. We turned out to be a good match. He wasn’t interested in yard work. Since his children are grown and it’s just the two of us, we make do. If cooking and cleaing are important to the person you fall in love with, work something out – like having that person do it while you pay for occasional maid service and dinner out. You don’t have to fit someone else’s ideal of a wife.
At my house, a good breaststroke will get you down the hall and we are generally satisfied with take-out food, microwavable stuff, fresh food, sandwiches, pizza or other delivery and snacks. We get our own meals. I plan to get one of those Roomba vacs and eventually paint everything dust colored.
I laughed big at the thought of some of the things you say you are afraid of doing and then you are out there in the Everglades with alligators! I break out in a cold sweat at the thought. Your job does sound cool!
You have to be able to clean house to have a husband or boyfriend? I can’t clean worth anything, and Mr. Neville and I get along fine in the mess we call home, with a little help from our housecleaner who comes by every two weeks. Mr. Neville’s a better cook than me, too.
Hoo boy! It’s like looking in a mirror. Believe me, we all screw up, sometimes costing hundreds or even thousands of dollars. I inadvertently made films that were the wrong size for big embossing dies, simply because I didn’t realize at the time that one of the settings in Acrobat was wrong. That was around $5000 dollars worth of error. Luckily, the bosses know I’m not stupid all the time, and forgave me.
All my life I’ve felt like “I’m smart. I understand things and I’m generally a quick study, so why do I screw up so much? Why can’t I get things together?”
I just have to remind myself, that nobody is perfect and we all have bad days, bad weeks, even bad months (don’t ask me about October from 1986-1996).
Just don’t let it get you down and keep plugging away. I concur with the others; your posts indicate an intelligent, thoughtful person who is perfect normal. At least, as normal as a Doper can be.
I’ll say what lots of people have been posting, that you sound so much like me. I had a little melt-down several years ago after making a mistake at work. A coworker overreacted and it felt like the end of the world. I spent a few hours telling my family how worthless I was. I even dredged up some big mistakes from the past so that I could use them to prove my case. I really did believe that I was so awkard it made me different from the rest of the world.
Cognitive therapy helped. “The Feel Good Handbook” is good. Sometimes I still feel big time insecure. Mostly though I realize that everyone makes mistakes and that having “book smarts” while being quiet and senstive (and a bit goofy) makes me different than most people, but in a good way. Apparently though, there are several of us here in this situation.
Oh, another thing I started doing was talking back to my mother when she told people, “NTBQ has no common sense” or "NTBQ has no sense of direction. " Or when she said, “You are too smart for your own good, NTBQ.”
I don’t allow myself to beat myself up over mistakes anymore, especially if the mistake involves somthing silly I did while trying to be more social. At least I tried to push myself to have fun and talk to people. Dwelling on mistakes only makes them seem worse. Of course, that doesn’t mean not learning from mistakes. I think you were very brave to go talk to the doctor about the mess-up.
And Finally… I swear I am not just saying this to be a sycophant …when I saw your post I thought, “Oh cool, that interesting girl, mostro wrote something in the Pit.” Honest.
