Life Science Tools of the Trade

keeps getting cheaper and cheaper.  We spend a LOT of time and money on DNA sequencing in our lab.  There is a core facility one building over and the convenience is great.  Bring your samples down (some DNA and primers in a tube) and two days later your sequencing data is sent via email.  Simple and easy enough?

Well, it just got easier.  Not only will they do your sequencing for you but, for about a dollar more, they’ll do the mini-preps for you as well.  All you need to do is bring some bacteria down.  The cost?  $3! 

From bacteria to sequencing data…. it doesn’t get much better.  At least, that’s what the grad student who is doing a screen keeps telling me.

Against my better judgement I listened to the rap of a green sales rep from Integrated Instrument Services. “We offer on-site pipette service at a great price…blah blah…freakin’ blah.”  It was pretty obvious he had memorized a routine, probably had no actual lab experience and couldn’t give a local reference (other than to say some people in some building on campus) use us.  For some unknown (idiotic) reason we decided to give this company a chance. As I posted before, we have been burned several times by unskilled monkeys who fix your pipettes with washers and springs from a hardware store.

Early the following  week a “technician” (not the hit and run sales dude) called and left a message. Unfortunately he never called back to set up an appointment. Late Thursday afternoon I called a pleasant  and apologetic young lady at the 1-800 number and complained. She acted truly concerned, and told me she would call the technician and he would be in the lab within minutes since he in fact was on campus but had simply forgotten us! True to her word he showed up several minutes later. I asked him if he had just received a phone call? He looked me in the eye and point-blank lied. “No, I did not get a phone call”. I explained that I had called the head office and was told you would come and offer a substantial discount.  He knew nothing about this and I guess it was simply a late Easter miracle that brought him to our door? I showed him the door and then called the corporate office back. I asked to be taken off all call and mail lists and asked that the sales rep not visit the lab again. I could deal with an honest “I’m so sorry, somehow I forgot to call back” but when the joker looks me in the eye and lies there was no way I was trusting them with this important job.

If a company offers such poor service when they are trying to gain new customers why would I deal with them in the future? Dirty details: The company is Integrated Instrument Services, corporate office headquartered in Indianapolis, IN. They may well do fine work in other areas of the country but I’ll never know. If you are located in the Carolinas or surrounding states I would avoid dealing with them at all costs.

Everytime I walked down the hallway to the lab, I kept seeing the ad for the new HaloTag system.  Well, finally, it got the better of me.  At the last vendor show, I spoke with the company sales rep about it.  She was helpful and provided me with some literature and one of the plasmids for cloning.  Thanks! 

If you haven’t heard of this system, the simplistic explanation is this:  you make a fusion protein of your gene of interest to an engineered derivative of the hydrolase gene.   Now that your protein is tagged, you express it in mammalian cells and then purchase differently labelled ligands to visualize it. 

What I’m not sure of… what’s the advantage is to this system?

The ligand covalently binds to the Halotag and comes in many flavors.  You’ve got your Biotin ligand, your coumarin ligand, your FITC ligand… Are you getting the picture?  You need to buy a different ligand depending on your experiment.  Presumably, how they plan on making money.

I was most interested in the immunofluorescence technology.  If it was going to save me time and frustration, I was all about changing over.  However, after reading a bit more, I didn’t see an advantage over traditional IF.  After fixing the cells, you bind the FITC ligand to your fusion protein and visual it with the scope.  How is that different than fixing cells and incubating with a FITC/TRITC conjugated antibody? 

The bottom line.  Either I’m missing the point of this technology or putting any tag on your protein will get you to the same spot.  I don’t see what a hydrolase fusion gets you over a myc or flag or HA tag. 

If anyone knows of a true advantage to this system or has actually used it, please let me know.  Until then, I’ll keep using my HA fusions for my experiments.

and, boy, it was a struggle.  After spending a week relaxing on a boat in the Caribbean, I had hoped to come back to work motivated and ready to go.  Not so.  It’s taken another week to remember what I was doing in the lab before I left and now, I remember why I took the vacation in the first place! 

Unfortunately, I’m almost to that point where it doesn’t even seem like I had time off…

I couldn’t help but feel a little uneasy when I heard the sales rep explain that the glove samples were “spritzed with glycerol”! Was that “for my pleasure” or ”for her pleasure”? I didn’t ask but it was way too close to a condom advertisement. The attractive disease-like pale green color was another real plus! Out of morbid curiosity, I tried on the sample glove which did not really fit like a glove. Verdict: Sorry, I don’t need lubricated colored gloves with reservoir tips.

It seems like not a month goes by when some guy with a flyer from some no-name company, a balance (maybe) and some rubber o-rings stops by the lab with an offer to “calibrate and repair” the pipetmen at ”a very competitive price”. Like the oil cartel and airlines, the prices are always in a very narrow range but unfortunately, the quality is not. I’ve seen a joker try and use a spring from a ballpoint pen and gaskets from Home Depot. I had the best luck with the Ranin repair woman but they haven’t returned several recent calls for quotes/service.

_____________ (www.annoyingspamcompany.com) is your best partner to outsource all those construct building works. With our large scale DNA sequencing facility, oligo synthesis capacity and experienced molecular biology research team, we can provide many services for a very competitive price. Our services include the followings:

1) PCR Cloning/Subcloning
2) Mutagenesis
3) Gene Manipulation (deletion, inserting, re-arrangement etc)
4) Construct building for fusion protein expression
5) De novo Gene Synthesis to obtain any gene construct ($1.25/bp).
6) Hairpin or other special construct building
7) ORF Cloning.
DNA purification and preparation.
9) Clone validation using sequencing or RE.

 Question: If I outsource this then what do I do? I don’t need a partner.
 

• iPod Shuffle
• 4-way Digital Timer
• Soccer Ball
• USB Flash Memory Stick

If those fabulous prizes caught your eye continue to read on. Otherwise, find something else to read.

Promega’s World Cup promotion: Buy three of our products, send in your World Cup card with your name and address. You (very probably) won’t win one of the four prizes but we’ll now have a huge mailing list to sell to other vendors! Get ready for some science SPAM! Call me cynical, but a chance to win an outdated iPOD, soccer ball with free air, USB memory stick or a  lab timer isn’t going to convince me to give any vendor my email and phone number.

Penalty kick!