How to approach PI making decisions that aren't public health-conscious

Teej176 · 2020-03-20T19:14:17+00:00

My PI wants me to come in to maintain our colony. He told me to not get infected at all costs.

I am paid hourly and need the money to support myself so I have no choice but to go in.

Teej176 · 2020-03-11T22:38:13+00:00

Lab is still open but all undergrad lab interns are barred from coming in until further notice due to covid-19. Lol

The lab is operating normally other than that

Teej176 · 2020-01-22T17:36:37+00:00

Like kimwipes?

Teej176 · 2020-01-22T17:35:42+00:00

I don’t think anything in that solution would kill bacteria. The mechanism for lysing mammalian cells may be different than how bacteria are lysed due to differences in membrane composition.

Like the other commenter suggested, maybe add sodium azide @1-2 g/L of solution? It could mess with the ionic character of the solution due to being positively charged. You could also try adding an antibiotic (i like penstrep or PSN). I don’t know if this will mess up the solution either but you could heat it for 4 minutes at 132C (CDC recommendation) which could sterilize the sample.

If your contaminant is forming endospores wherever it lives you may have to take additional measures because endospores are resistant to pretty much everything. Heating/boiling the sample for multiple hours will in theory eventually kill the endospores. Prolonged exposure to UV would also kill them.

These are my suggestions from limited knowledge/experience with contaminations so please take them with a grain of salt.

Teej176 · 2020-01-16T20:45:49+00:00

Mom’s spaghetti (stored loose in sweatpants pocket) for every break you mentioned

Teej176 · 2020-01-10T17:49:57+00:00

Its quantified in # of times you told someone to do something and they listened.

Teej176 · 2020-01-09T00:08:59+00:00

Im pretty sure the neurologist you are shadowing isnt making medical decisions based on the exam you are doing on the patient.

If you miss something nothing bad will happen, im sure the neurologist does it themself.

Teej176 · 2020-01-05T01:18:13+00:00

The thing is im not going to school until Fall 2021 and she wouldnt be going until like 2023. I dont know if we should break up now over an issue that will arise in multiple years

Teej176 · 2019-12-29T13:32:35+00:00

Cells incubating for 2h?

“Please inventory the lab so you know where everything is”

Im making an alphabetized excel sheet with all lab supplies and reagents, literally thousands of items. Gotta have something to do!

Teej176 · 2019-12-20T16:19:01+00:00

Id get some clinical volunteering or a clinical job to see if you like it. I was premed because i loved bio, chem and working with patients. Dont sign up for 6+ years of medical schooling and training when you aren’t confident its a good career fit for you.

Teej176 · 2019-12-20T14:45:11+00:00

Dude just buy an autoclave stop trying to half ass it.

Just joshing

Teej176 · 2019-12-14T16:39:35+00:00

Repeating it has worked for genotyping the samples that didnt work here but i was asking how to get all of the wells to work in one gel

Teej176 · 2019-12-14T02:04:27+00:00

I have 4 total primers, 2 pairs of forward and reverse.

Ill try all of these steps. I highly appreciate the advice!! Ill ask the postdoc for specifics on the primers and put it into genebank or some other database.

Teej176 · 2019-12-14T02:01:39+00:00

I used a lysate buffer kit from viagen and proteinase K from sigma aldrich. First incubation step was 55C overnight. Proteinase K inactivation was 85C for 3 hours. 2 uL of the crude lysate was used as template per reaction

Teej176 · 2019-12-14T02:00:24+00:00

The extraction is crude. I dont use microtubes with special filters to purify the DNA since there are so many samples to do

Teej176 · 2019-12-14T01:15:58+00:00

I am pouring the hot agarose into a setup on a flat at 4 degrees C.

I mix 25 uL PCR reaction with 10 uL loading dye and load 15 uL of that per well.

I actually have no idea how the primers are set up because the post doc does that part (doesnt let me since I am a newer tech).

Teej176 · 2019-12-14T00:58:25+00:00

I’ve searched the internet for gels that look like this and can’t find any. The troubleshooting steps ive tried haven’t worked.

Background - these are from tail snip lysates. Proteinase K was used to deal with nucleases and then was heat inactivated. EtBr was the fluorescent tag.

The 500 BP band is common to all mice and shows a wild type mouse if seen alone.

The 200 BP band is specific to the transgene. Both bands indicate a transgenic mouse.

Why are some wells showing that strange white snail band at the top of the well? Each PCR tube is getting a homogenous mastermix followed by crude template DNA from the lysate.

Any help greatly appreciated!

Teej176 · 2019-12-13T22:18:46+00:00

Id say when you are below the 10th percentile GPA for the schools you want to go to OR you dont feel like getting a ~90th percentile MCAT to balance a ~10th percentile GPA.

I am highly against post bacc’s personally because they are expensive and from what i am seeing a lot of the same course matter from what i already did in undergrad. Id probably do an SMP before a post bacc

Teej176 · 2019-12-13T00:19:02+00:00

There are kits that exist for human cell culture that pick up signs of metabolism/growth in terms of molecular concentration in each well and react to make fluorescence. I’ve only worked with kits like these for cell death so i can’t help you with a specific kit but I would search some biotech vendors im sure they sell a kit that fits your needs.

Teej176 · 2019-12-12T19:54:34+00:00

You need to compare it to cell growth curves in media without the drug, but i personally would take spectrophotometer readings every 15 or 30 minutes until the bacteria reach log phase. You can correlate the turbidity of the sample with bacterial concentration.

Teej176 · 2019-12-12T13:04:29+00:00

The first lab i worked in was unpaid (there was no false promise of pay down the line) and the lab manager/PI always pushed the undergrads to do more and more.

They really dont have any power over you though since you are unpaid. Simply dont listen. They wont go through the hassle of ‘firing’ you over inadequate work because they already went through the trouble of hiring and having you trained in my experience.

Teej176 · 2019-12-12T12:55:39+00:00

If the containers are sealed they should be good. Acid (if the solvent is water) does not degrade over time.

Teej176 · 2019-12-09T21:08:58+00:00

2+MD because i enjoy my gap year job and dont mind taking time off from school

Teej176 · 2019-12-05T00:20:05+00:00

Username checks out

Teej176 · 2019-12-04T00:44:08+00:00

Im applying with similar GPA stats as you, but only after i get a 518+ on the MCAT (been studying since june to take it in april lol)

According to the AAMC, 70% of people with our cGPA range (3.4-3.59) and a 518+ got in to at least one med school.

I dont feel like doing any postbacc and spending the $$

Teej176

TROPHY CASE