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Author: Brian Krueger, PhD | Views: 16254 | Comments: 0

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If you’re an avid follower of popular science in today’s news media, you might have noticed a recurring theme. Genomics is everywhere. On an almost weekly basis, the New York Times, the New Yorker, Forbes and a myriad of other outlets are publishing stories with overly optimistic ledes about doctors and gene sequencers being replaced by apps and iPhone accessories. You would be forgiven if you thought genomics was “solved” and we’re 5 years out from creating a Star Trek inspired “tricorder” that near instantly sequences your genome and tells you, without equivocation, what malady is afflicting you and how exactly to overcome said disorder. The fact of the matter is that we’re not there yet, not by a long shot.

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Author: Brian Krueger, PhD | Views: 6382 | Comments: 0
The Advances in Genome Biotechnology conference starts tomorrow in Marco Island, FL. Twitter and the blogs have been a flurry of speculation about what the major vendors will present at this years’ meeting. In previous years we’ve seen the introduction of new, “disruptive” technologies such as the ion torrent platform, the Oxford Nanopore Minion and the PacBio RS. Like many, I have mixed emotions about this conference. It’s more CES than science. Given the history of the major announcements and where those products are now 3 and 5 years out it’s hard to get excited about a show stopper. While technically impressive, the MinIon is still mired in problems that were glossed over in the fanfare of the original announcement and PacBio is FINALLY starting to deliver on the promises it made eons ago. I should also mention my disappointment with Ion Torrent here. This is yet another company that made a major announcement at AGBT and failed spectacularly. Keith Robison thinks they still have a sho . . . More
Author: Brian Krueger, PhD | Views: 6772 | Comments: 0
Yesterday marked the kickoff of the JP Morgan Healthcare Conference in San Francisco. Following last years’ lead, Illumina once again used this platform as an opportunity to announce the release of a number of new products including 4 “New” sequencing systems ahead of the more scientifically focused Advances in Genome Biotechnology conference in February. At this same time in 2014, Illumina presented the HiSeq X ten sequencing system which is a system for population scale genomics composed of ten HiSeq X sequencers. Illumina touted this system’s reduced reagent price, increased speed, and expanded capacity. It has now taken much of the technology from this HiSeq X system and put it into two new lower tier models: the HiSeq 3000 and HiSeq 4000.

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Author: Brian Krueger, PhD | Views: 6183 | Comments: 0
Illumina, the world leader in short read DNA sequencing, made a series of very big announcements yesterday at the JPMorgan Healthcare Conference. These developments have many sequencing labs around the world excited and worried all at the same time. The excitement comes from the fact that it appears on the surface that Illumina has broken the $1000 genome* barrier – the worry comes from the realization that only a few of us can afford it.

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Author: Brian Krueger, PhD | Views: 17356 | Comments: 9
Last by Michael Schatz on Feb 26, 2013, 12:13am
Aside from the dubstep pumping out of the Roche and Agilent booths, the volume of AGBT has been somewhat muted. There was no grand offering of new hardware or over the top promises of sequencing genomes on what now appear to be vaporware USB thumb drives. This is my first in person experience of AGBT, so as a virgin it seems for the most part to be rooted in the science despite the ridiculous parties and “showgirl” casino nights. The atmosphere here is unlike any other science conference I’ve attended. It’s like the bastard child of a Gordon Conference and a Las Vegas Porn Convention. I really hope that the deep pockets of Sequencing Centers are more influenced by the science than the free dinner parties and alcohol, but I have pretty low confidence in humanity. Regardless, I think everyone in attendance today was overwhelmed by a stunning talk from PacBio and the dramatic advancements of their long read technology.

The PacBio talk came on the heels of what felt like a warm-up opening act from Jeremy Schmutz of the Hudson Alpha Institute. Schmutz has been working with a start-up that was recently acquired by illumina called . . . More
Author: Brian Krueger, PhD | Views: 7335 | Comments: 0
As the second day of AGBT kicked off, it became quite clear that this meeting would be dominated by medical genomics. There were a few talks sprinkled in about gut or sewer microbiomes but the vast majority of the talks the last two days have been on clinical genomic sequencing. This is fine by me since it’s exactly what we do in the Genomic Analysis Facility in Duke’s Center for Human Genome Variation. It’s really nice to see how other centers are approaching these problems. Unfortunately, this is one of the few opportunities we have to peek into each other’s operations.

Yesterday’s first talk was by Russ Altman of Stanford University. Russ has been a leader in the field of pharmacogenomics and he presented his work on developing the Pharmacogenomics Knowledgebase (PharmGKB, pharmGKB.org). He led by saying, “Don’t ever give a talk about a website,” and in his case it was true because WiFi in the conference room was down for the majority of his talk. He urged the crowd to follow along on the website, but only those of us with a cell connection could join him. Russ pointed out the major drawbacks of using GWAS and SNP chips for obtaining information about pharmacoenomic associations and joined pretty much everyone else in saying that the standard t . . . More
Author: Brian Krueger, PhD | Views: 7224 | Comments: 2
Last by Brian Krueger, PhD on Feb 21, 2013, 11:42pm
Today I dusted off my luggage and headed down to the annual Advances in Genome Biology and Technology meeting in Marco Island, FL.  Historically, this meeting has been the Detroit Auto Show of Genomics where companies and labs release their beautiful shiny new products and methodologies.  In past years, attendees were showered with fireworks displays and epic swag bags.  The tone this year is palpably more mutated.  One only has to point to the display banners located on the AGBT presenter stage for evidence of this: banners for Bronze and Silver Sponsors appear to hang waiting for accompaniment by Gold and Platinum brethren…there’s even space allocated for them.  

Despite the tightened biotech purse strings, the event appears from the outset to be extremely well organized.  There could be a few more power outlets for those of us carrying a small fortune in lithium powered devices, but I guess I’ll manage.  And of course there’s no live streaming or any form of web enabled anything…but I can gripe about that in a long winded and whiny post next week.

The evening opened with a brief introduction with a description of some new meeting changes.  This is the biggest AGBT yet with over 800 attendees.  A new abstract selection committee was created . . . More
Author: Brian Krueger, PhD | Views: 4245 | Comments: 3
Last by Brian Krueger, PhD on Sep 28, 2012, 5:02pm
One of the big stories that blew up on the internet the other day was the publication of some results that reportedly show that females who have birthed males have male DNA in their brain. That’s pretty cool stuff! This isn’t uncommon, it’s called microchimerism, or the deposition of cells or DNA from the fetus to the mother or vice versa. It has been shown in both mice and humans that the blood of the mother contains snippets of fetal DNA and even whole cells. These have even been used to completely sequence a fetal genome using only the blood of the mother. The finding that these DNA fragments or cells can deposit themselves in the human brain is novel, and potentially cool from a brain regulation standpoint. Maybe baby boy brain cells change the mother’s brain?

However, a quick read of the abstract of the paper yesterday immediately raised a few red flags in my own brain (absolutely full of male cells, by the way). The “discovery” was made using quantitative real-time PCR and only quantitative real-time PCR on a highly repetitive male gene on the Y chromosome. Most people who aren’t scientists don’t . . . More
Author: Brian Krueger, PhD | Views: 4976 | Comments: 2
Last by Brian Krueger, PhD on Sep 08, 2012, 12:05pm
In 1990, the scientific community embarked on a landmark experiment to completely sequence the human genome. At the time, it was assumed that knowing the exact sequence of the human genome would provide scientists all of the information they ever wanted to know about genomics and how DNA contributes to human disease. At least this is how the project was presented to the public, however, every genome scientist knew that obtaining the sequence of the human genome was a lot like getting a cake recipe that listed all of the ingredients but didn't explain at all how much of each ingredient to use, how to mix the ingredients together or how long to bake that mixture to create a delicious cake. Compound that with the fact that the list contains over 35,000 ingredients and you can quickly understand why the sequence alone wasn’t very informative. We spent over 3 billion dollars to obtain this sequence and since the final publication of the sequence in 2003, the media has questioned its usefulness. Apparently, because cancer wasn’t cured in 10 years, the human genome project is largely seen by the media as a colossal waste of money; however, obtaining this sequence has been invaluable in speeding up research and has significantly contributed to our understanding of how som . . . More
Author: Brian Krueger, PhD | Views: 3141 | Comments: 0
In the current political climate it has become clear that science is a major target of Republican directed budget cuts. However, the soundbytes of politics do not represent the importance of science in our lives. Because of this, I think it's extremely important that we explain why some of our model systems are so important for understanding how viruses and ultimately human disease work.

In the lab that I run, we currently work on mutating two different herpesviruses. One of these is Kaposi's Sarcoma Herpesvirus (KSHV) and the other is Murid Herpesvirus 68 (MHV68). Both of these viruses are gammaherpesviruses. In humans, KSHV only really ever becomes a problem in individuals who have a compromised immune system such as those infected with human immunodeficiency virus (HIV). KSHV is an interesting virus because its default program is latency, meaning that once it gets into your cells, it turns itself off and waits for conditions which allow it to grow and take over. This is akin to a bear hibernating in the winter. We do not understand how or w . . . More
Author: Brian Krueger, PhD | Views: 687 | Comments: 7
Last by FunkDoctorX on Aug 03, 2011, 7:27pm
A week or so ago someone forgot to close the door on our enzyme freezer tightly. I had just ordered $1,000 worth of NEB enzymes to make high throughput sequencing libraries too... Before the meltdown, I made a couple of test libraries to be sure that the protocol was worked out.


One of my test libraries with a perfect library smear. We extract the DNA in between the 200 and 300bp bands for sequencing.
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Author: Angry Scientist | Views: 8061 | Comments: 3
Last by Moderates_Rule on Mar 26, 2013, 11:56am


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Author: Brian Krueger, PhD | Views: 697 | Comments: 2
Last by Brian Krueger, PhD on Mar 17, 2011, 10:17am
Today I noticed my Tomato Clownfish acting a little friskier than usual. A few days ago I saw a white bump under the female's anal fin and thought it might be a fungal infection because it had a goofy gray tinge to it. I figured I'd just wait and see what developed. Well today I noticed that it was much bigger and longer...It was her ovipositor (egg laying tube)!! She also had a big fat round belly, so that really means only one thing. I watched her and the male clownfish do their dance and the ovipositor grew longer while the male started showing signs of arousal too. I guess a video is worth more than my explanation. I started recording after the first egg went down. It's the little orange sac in the middle of the screen. This patch grows much larger over the course of the 10 minute video. Enjoy!

Mass eggs start going down around the 2 minute mark if you don't want to watch the fish dance.

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Author: Brian Krueger, PhD | Views: 5178 | Comments: 8
Last by americanbiotech on Mar 10, 2011, 12:22pm
Every once in a while I find myself having a "grass is greener" moment in science. I sit at the computer thinking about all of the annoying things in lab that aren't working and wonder, "If I could answer any question, given unlimited resources, what would I choose to study?" This, my friends, is where I prove my unhealthy obsession with fish.

If money, fame, and science groupies weren't a priority, I'd spend my time and resources trying to find a cure for Cryptocaryon irritans, which is better known in the tropical fish industry as white spot disease, Ich, or Crypto. Crypto is a nasty little bug. It's a protozoan ectoparasite that lodges itself in the epidermis of fish where it grows until it drops off to mature into an Aliens inspired cyst that spits out 300 new little bastard parasites that go on to infect more host fish. In the open ocean, this guy isn't so terrible because its chances of infecting a host are minimal considering the massive amount of water a parasite has to travel through to find a new host. This is a completely different story at an aquaculture facility or in the home aquarium where fish are typically stocked to capacity in relatively small volumes of water. A single cyst can quickly turn into a fish killing epidemic. . . . More
Author: Brian Krueger, PhD | Views: 543 | Comments: 1
Last by Dr. Girlfriend on Mar 02, 2011, 2:11pm
Things have started to calm down a bit on my weekends so I've had more time to attend to fish. One of the crappiest things about owning a saltwater fish tank is that you can't just add fish to your system immediately. The fish come from the ocean covered in parasites, so you have to take extra special care of new additions to be sure they're clear of parasites and infections before you add them to your main "display" tank. Quarantining fish is absolutely a necessary evil. A lot of people neglect to do this, but it saves a ton of headaches. Trust me on this, you would much rather lose an $80 fish than get your $3000 display tank infested with a parasite!

Anyway, the fall got clogged with trips and holidays. Some of you may remember that back in October I tried to quarantine a yellow longnose butterfly fish and a powder blue tang that both died from a nasty bacterial infection. Given that quarantining fish takes at most 2 months if they get sick, I didn't have enough time to cycle in more fish before Turkey Day! Once Christmas passed, I was working on setting up an office tank for Whitney, so my quarantine system has been occupied with her inhabitants for the past two months... Finally I was able to pick up a new butterfly fish a few weeks ago. . . . More
Author: Brian Krueger, PhD | Views: 2405 | Comments: 11
Last by Thomas Joseph on Jan 04, 2011, 7:47am
I'm beginning a new project in lab. It's a series of ChIP-seq experiments and the first step to doing ChIP-seq properly is optimizing sonication conditions. Here's a trial run with the sonicator I plan on using. The DNA shown in the gel is from cells containing latent herpes virus. We're looking to shear the DNA so that the bulk of it is between 100 and 600bp. For ChIP-seq we extract and purify the DNA in the 100-300 range in the gel. Looks like about 13 cycles of sonication should do (sonication past this point doesn't result in smaller fragments, don't want to risk over sonicating)! Actually, I think I'm going to do this again on Monday. I looked back at some old data and I should be able to get the fragment sizes a bit smaller by increasing the pulse time. I'll post an update soon!



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Author: Brian Krueger, PhD | Views: 6182 | Comments: 1
Last by Evie on Dec 18, 2010, 11:17am
Yesterday there was some buzz over at Huffington Post about a stem cell cure for HIV. I first ran across the article via a link a friend of mine had posted on Facebook. The HuffPo piece is scant on details, so I’ll provide a quick run down on what’s going on here. But first, a lesson in HIV virology…

Human immunodeficiency virus (HIV) was first discovered in the 1980’s when gay men and IV drug users started turning up in hospitals with very odd opportunistic infections like Kaposi’s Sarcoma Herpes virus. These individuals had severely compromised immune systems and the original name given to the condition was gay related immunodeficiency disorder (GRID). The discovery of a viral cause of the disease came in 1983 from the labs of Luc Montagnier (recently won the Nobel Prize for this work) and Robert Gallo (recently didn’t win the Nobel Prize and is kind of pissed about it).

Genetic tests have shown that HIV originated in African monkeys and is related to a similar condition in monkeys called Simian Immunodeficiency Virus (SIV). It is thought that the virus was passed on to humans through the consumption of “bush meat” in sub-saharan . . . More
Author: Brian Krueger, PhD | Views: 2534 | Comments: 3
Last by Evie on Dec 18, 2010, 10:48am
In a previous post I mentioned that I spent the first year of graduate school working on a dead end project that was going nowhere. I’m going to present and discuss the findings of the paper that changed the direction of my thesis project and ultimately lead to the completion of my PhD.

In the fall of 2006, I was finishing up the last few experiments on a project trying to link P53 (a very important protein in cells that helps to prevent cancer) to one of the most important transcriptional activators, P-TEFb. I should probably pause for a second here to give a little bit of background on P-TEFb so that what follows is somewhat understandable. Positive transcription elongation factor b (P-TEFb: Pee Tef bee) is a protein kinase (read as on/off switch) that regulates RNA polymerase II transcription of most genes. RNA polymerase II is the protein responsible for messenger RNA (mRNA) transcription or the process of turning DNA into mRNA. Without mRNA, the transfer form of DNA, proteins cannot be made by the cell, and life does not exist. Essentially, P-TEFb tells RNA polymerase when it’s ok to start making RNA. You can think of P-TEFb as the starter at a track meet, you know, the guy th . . . More
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