A Long Overdue Update

My apologies for taking so long to post anything new. Between medical school applications and research, the past few weeks have been pretty busy. I'll try to summarize what we've been up to below, mostly in the form of pictures.

Week 2:
Dr. Hamilton wanted to run gels to compare the proteins in the different beer samples. Before doing so, I ran a Bradford assay to make sure I didn't overload the gels.

Preparation of cuvettes for Bradford.

Spec and cuvettes ready for analysis.

Starting polymerization on 10% SDS gels

Gels running

Gels just before destain

Gel #1

Gel #2

Unexpectedly, the Bradford assay returned protein concentrations relatively equal across all samples, and the gels showed the same bands with the same intensities throughout, the largest of which was just over ~35 kD.


One of the required reagents for the RIDASCREEN ELISA is a solution of ethanol and fish gelatin. This reagent is a pain to make as the ethanol can cause the fish gelatin to crash out of solution. The gelatin is also about twice the viscosity of honey. Below is a picture of me trying to vortex the mixture to get everything into solution.

Fish gelatin and ethanol!

All 12 gallons. Notice how the 1 step (on the right) have a lot more trub than the 3 step.

Full RIDASCREEN Plate:
Below are pictures of the RIDASCREEN ELISA performed on a full plate (verses the semi-full plate shown before)

Prepping of samples. 1 mL of beer + 9 mL of fish gelatin/ethanol solution.

Supernatant and a diluent post centrifugation and ready to add.

Standards and samples added along with the enzyme conjugate.

Incubating on the rocker.

Samples post-incubation.

Plate after third wash with 250 µL wash buffer.

Plate after addition of 100 µL of chromagen to each well.

Plate after incubating with chromagen for 10 minutes.

After addition of 100 µL stop reagent to each well. Read in the plate reader immediately.

My best friend for the ELISA. It allows me to pipette the same amount multiple times without refilling or changing the tip. It speeds up the process of having to pipette into all 96 wells.

After our third assay (through Day 10 of fermentation), we determined that all of the samples with Clarity Ferm added were almost completely gluten free, all reading less than 4 ppm (<20 ppm can be sold as being gluten free). We will continue to take samples and analyze them after bottling to make sure that the beer truly is gluten free.

Week 3:
Trip to Ann Arbor
Dr. Hamilton and I needed to make a trip to Ann Arbor to pick up some brewing supplies at Adventures in Home Brewing. Since we were already there, we decided to stop at Arbor Brewing for lunch. The beer was definitely worth the detour.

Flight of everything Arbor Brewing had on tap.

Dr. Hamilton enjoying some of the flight.

Bottling!

Dr. Hamilton taking a minute to help scrub some bottles. It took a lot longer than we thought to get all the bottles we would need clean and ready to go. Thankfully, we started the day before bottling.

About half of the bottles we needed. Thanks to Dr. VanZant for giving us extra bottles!

After struggling to get the bottles upside down to dry, I decided to turn the cooler on its side to stack the bottles. To my surprise, it worked perfectly. The bottles were left upside down overnight to drain excess PBW.

In the midst of bottling, a new UV-Vis spectrophotometer came! I felt like a kid on Christmas morning helping set it up.

Prepped and ready to start bottling.

Sanitizing a bottle before adding the beer.

 

Another view of the sanitizing.

 

Racking from the carboy to bottling bucket with priming sugar added (18 g sugar/gallon of beer if you were wondering).

  

Dr. Hamilton just finished bottling in this picture.

For some reason, the capper was super cool to me.

All of our bottles after two mornings of bottling.

I get really defensive of my beer, er, research.

What kind of people would we be if we wasted the extra? Even before carbonation, it tastes pretty good.

Milling:
Since the efficiency of the stout we brewed was a little low, we decided to mill the grains for our Oberon clone a bit more than the company we bought them from online had. We'll be brewing this on Monday and Tuesday.

Milling the grain for the Oberon clone.

Power tools are absolutely fantastic. Dr. Hamilton also knows how to capture me at my finest.

Check back for updates periodically! After a few weeks we'll be able to test the stout for taste and final gluten content. I already have a list of people willing to help, for science of course.

Bonus picture of my housemate Wyatt celebrating that his research on MRSA seems to be working