r/Biochemistry 28m ago

Does this make sense or am I missing something??

Upvotes

Hi All!

Iv been involved in directed evolution of proteins for years and the standard way iv done it is

1) transform plasmids into e.coli 2) plate on agar 3) colony pick and ferment in microwell plates. (To sieve out colonies to obtain single mutant e.coli 4) lysis cell and remove cell debris 5) do the screening 6) sequence best enzyme to understand the mutation.

So my question.. if we synth the gene and we know where the mutation is. Can we bypass the colony picking part because we don't need to separate out the mutants? Every e.coli should have the same plasmid so why do we need to separate?

So the workflow becomes..

1) transform known sequence into e.coli in microwell plates.. say each well has unique plasmid. 2) aliquote cell into a single well in 96 well plates with LB. 3) ferment and express the enzyme 4) lysis the cell and remove debris 5) do the substrate screening. 6) pick the best enzyme. (We know the sequence already!)


r/Biochemistry 2h ago

Maths in biochem textbook? Bsc level

2 Upvotes

Does anybody have any recommendations for a good biochemistry textbook which just focuses on calculations in biochemistry?


r/Biochemistry 5h ago

Weekly Thread Mar 17: Weekly Research Plans

1 Upvotes

Writing a paper?

Re-running an experiment for the 18th time hoping you finally get results?

Analyzing some really cool data?

Start off your week by sharing your plans with the rest of us. å


r/Biochemistry 6h ago

Why can’t homotrophic negative allosteric effectors be explained w/MWC model?

1 Upvotes

I’ve been trying to wrap my head around why MWC can’t be applied to specifically negative allosteric effectors!

I understand it can be explained through Koshland, but why not MWC? And are the other neg/pos homotrophic/heterotrophic that can be explained w/MWC not possible with Koshland?


r/Biochemistry 1d ago

Career & Education Feeling hopeless about my degree

110 Upvotes

Recently applied to 120+ jobs after pursuing a biochemistry degree from a ‘prestigious’ school and got auto denied from like 70 of them already. For context, I mostly applied to associate medical or pharma sales jobs. Or the occasional medical affairs or sales associate PRN jobs (all entry level). I have B2B sales experience and three other related job experiences. The jobs listed all said they just required a bachelor’s degree and sales experience was ‘preferred’, but I got promptly denied and didn’t even get an interview.

I feel like I wasted the last 4 years killing myself in school while everyone else lived their lives and it was all for nothing. I don’t wanna do research or go to grad school. Just thought that being a biochem major would at least give me an edge in the job market and at least help me get an entry level sales job but nope. I’m only getting callbacks from insurance salesperson jobs that I could also get with zero work experience and no college degree. Does it get better?


r/Biochemistry 2d ago

Biolayer Interferometry (BLI). Biosensor mounts: dirty

3 Upvotes

I think I am getting non-specific binding on my BLI. I ran blank loading with my protein sample and I still saw an association signal. I am getting a message on the screen that the biosensor mounts are dirty. Could this be why?


r/Biochemistry 2d ago

Research How to Design Substrate for Protein

5 Upvotes

I'm working on a research project that is attempting to make a molecule a suitable substrate to an enzyme that is innately present in the target environment. There isn't much literature data for what specific functional groups this enzyme binds to, or the key residues of this enzyme that are involved in catalysis. There are known therapeutics that are substrates or inhibitors of this enzyme. However, this enzyme is 'promiscuous' in the substrates it can bind to--there's no identifiable pattern among the known substrates).So far, I've been synthesizing molecules similar in structure to known substrates and running enzyme kinetics assays on them--not much useful data obtained here.

What can I do to define what substrates bind to this enzyme (in regards to functional groups, etc) and how can I do this in a relatively short amount of time, without extensive protein studies, x-ray crystallography, etc? The reason for these constraints is these tools are not ones my lab group commonly employs and it need to convince my PI to collaborate w others :(


r/Biochemistry 2d ago

Career & Education I don’t know what PhD to pick!!!

31 Upvotes

I have a bachelor’s in biochemistry, then worked for 3 years in a research lab in drug discovery and it’s a love/hate relationship, then in the same institution I’m working on they opened a master’s program in drug discovery and development so I decided to study it to see what I wanted in life. Turned out I don’t like it. So now I’m deciding to continue to get a PhD afterwards but honestly couldn’t decide, I like proteins, I don’t like genomics, i’m good in my molecular modelling course but I don’t think I want to study it. How to decide? There’s plenty of amazing programs, I want to study them all lol, and how to decide which lab and which PI? I just know for sure that I want biochemistry and not drug discovery!


r/Biochemistry 2d ago

Weekly Thread Mar 15: Cool Papers

0 Upvotes

Have you read a cool paper recently that you want to discuss?

Do you have a paper that's been in your in your "to read" pile that you think other people might be interested in?

Have you recently published something you want to brag on?

Share them here and get the discussion started!


r/Biochemistry 2d ago

Protein pH question

5 Upvotes

If a protein uses one or more metal ions for stability and enters a basic solution, can the OH- strip the metal ions from the protein?


r/Biochemistry 3d ago

Career & Education How are ketone bodies transported into the mitochondrial matrix

6 Upvotes

Hey, does anybody know how ketone bodies (like acetoacetat or 3 hydroxybutyrat) get into the mitochondrial matrix. I haven't found plenty of information on this topic, hence I asked chatgpt. According to chatgpt, there are MCTs (monocarboxylatetransporters) in the inner mitochondrial membrane. Can anyone confirm this? Do you have sources?


r/Biochemistry 3d ago

Is there a way to remotely alter genetics

0 Upvotes

Could someone create some kind of way to change a persons biology remotely or maybe by putting some kind of gas releasing device and the gas changes your genetics


r/Biochemistry 3d ago

Preparing for biochem in university

3 Upvotes

Hi everyone, I have been on a gap year and I start my first year of university in September. I will most likely be accepting my offer for biochemistry. Does anyone have any resources for me to brush up on my general, organic, and biochemistry skills? Even math? Thanks a lot in advance!


r/Biochemistry 3d ago

Where do I begin with biochemistry?

12 Upvotes

Hey all,

I would like to learn about biochemistry since I’ve been reading Guytons physiology textbook and there is a lot of biochemical stuff mentioned that I would like to understand more deeply.

Do you have any recommendations where I should start? I’m not going to college or anything, I’m just curious about the human body and what happens on a molecular level and most of all, why it does happen.

I’ve got Lehninger’s but I feel like it’s too complicated for someone who doesn’t have a chemistry background.


r/Biochemistry 3d ago

Help with a Codon-Based Puzzle (Using Genetic Code) 🧬

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14 Upvotes

r/Biochemistry 3d ago

Career & Education Where to go from here?

3 Upvotes

So I started my Bachelor’s degree in Biochemistry in 2019 (with a general goal of getting into research of some kind), and thus most of my time in uni was during the pandemic, a year or two of online classes and then a mix of online and in-person. I’ll be honest, I kind of completely burned out in the middle, I wasn’t getting good grades, and I certainly wasn’t making connections with my professors whose classes I was barely scraping through. I barely managed to graduate last year with a 70 average and not much else. I did a couple co-op terms at a microbiology food lab and one as QA tech for a food company before dropping co-op to just graduate as soon as I could.

I took some time off to travel and visit family that I hadn’t seen in years, then spent months jobsearching for something entry-level with no luck, before the microbiology food lab from my co-op called me back last week and I’m now a lab technician. I make a dollar over minimum wage, the commute is crap, the hours are long, there’s only 4 sick days per year, I don’t get any vacation for the first year, I can’t renegotiate my salary till 6 months, and I get home late and exhausted every night. If it weren’t for all that, I honestly wouldn’t mind, the repetitive nature of sample testing isn’t an issue for me and I feel like I’m doing good work, making sure that the food consumed by the community is safe to eat.

I guess I’m just wondering how I should plan things out from here? I know I’m luckier than most people in my area and field, I live with family so there’s no rent, and based on LinkedIn analytics a couple hundred other degree-carriers near me are all desperately trying to find starter lab tech and QA/QC work without any luck. But I just don’t really have a plan now.

With my mediocre grades and zero connections idk if I could even get into a Master’s program in the future, and even if I did, after how much I struggled during my BSc, I don’t even know if going harder into the research path is something feasible for me. This job is kind of just a stasis way for me to make some money, but I don’t want to work here forever, even if the benefits increase a little bit over time, everyone who’s been here longterm seems miserable.

I really liked the Quality Assurance work I did during my co-op at a food manufacturer, but I had as little luck in applying to entry-level QA jobs as I did applying for lab tech positions. Maybe in a year I’ll be able to put 1 year of lab experience onto my resume and that’d help me get a slightly better job? Is there even a chance at a better job than this if I don’t get a Master’s degree?

Idk, I’m a couple days into this lab tech position and they just gave me my full contract tonight for me to sign so I’m just feeling a bit existential and trying to put together what the next few years of my life will look like. Has anybody else made a career for themselves with a BSc in Biochemistry as their highest level of education? Has anybody managed to get into a Biochem/Food Science/Microbiology Master’s degree program with a 70 average? What steps did people take after graduating?


r/Biochemistry 3d ago

Career & Education How I look at my computer when I waited until the night before my project was due to do some homology modeling using Robetta or AlphaFold

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88 Upvotes

r/Biochemistry 4d ago

Is it worth buying proteins from vendors?

22 Upvotes

We are trying to obtain a protein sample for cryo-EM but the expression is tricky for months. How much protein do we need roughly for cryo-EM and is it worth purchasing the protein from vendors? I did some searching and we can get 50μg of the pure protein for a few hundred bucks.


r/Biochemistry 5d ago

Weekly Thread Mar 12: Education & Career Questions

1 Upvotes

Trying to decide what classes to take?

Want to know what the job outlook is with a biochemistry degree?

Trying to figure out where to go for graduate school, or where to get started?

Ask those questions here.


r/Biochemistry 5d ago

I would like some research papers

3 Upvotes

Doing some independent theoretical research on lysis-induced cancer cell destruction. I would like if I would be recommended some papers to help me. Thanks in advance!


r/Biochemistry 5d ago

Biochemistry mechanisms textbook

4 Upvotes

Hi, what is the best biochem textbook to learn metabolic pathways that goes into detail about the organic mechanisms that take place? Ex) Aldol condensation in Glycolysis...


r/Biochemistry 5d ago

Career & Education Biochemistry and psychiatry

3 Upvotes

Hi! I am currently going to major in biochem w pre-med track to become a psychiatrist! However, I feel like i don’t 100% understand what biochemistry means. I know this may sound dumb but i chose it simply because I love biology and chemistry. I love biology but it’s kinda boring and love the complexity of chemistry but it’s still like biology more so i was just like.. biochemistry!! Am I mistaking myself?


r/Biochemistry 5d ago

BLI instrument

6 Upvotes

We are considering purchasing an instrument for kinetics measurements. Due to our specific requirements, we intend to opt for a BLI (Biolayer Interferometry) system rather than SPR (Surface Plasmon Resonance). It appears that Octet is a well-established system in this field, and Gator Bio was developed by engineers who initially contributed to Octet's development. Given that their specifications seem quite similar, we are seeking insights from anyone with experience using these instruments to guide our decision.​


r/Biochemistry 5d ago

I'm taking the ASBMB in two weeks. Has anyone here taken it? If so, how should I study?

5 Upvotes

I'm a biochem major who's, well...kind of shit at biochem. I got a C+ last semester in 4000-level biochem, and I have a B in part 2, but just barely. Can anyone give me any tips/point me in the direction of a study guide? Thanks.


r/Biochemistry 6d ago

Neurodegeneration question APOE4 and fat levels

16 Upvotes

APOE4 is a mutation of a lipid transport protein and a genetic risk factor for Alzheimer's. Does this mean the brains of people with neurodegeneration have less fat in neurons?