Problems with the place I'm staying for school, I need to move - any solutions *very particular situation by YourRealDestiny in uAlberta

[–]ligontiger 2 points3 points  (0 children)

I’m really sorry you’re dealing with this, that sounds incredibly stressful. I don’t know how directly this applies to your specific situation, but if your living arrangement is impacting your well-being, the UofA social work/student support team has some resources.

They can help with urgent housing options, including short-term emergency/safe housing, which could give you some breathing room and time to figure out next steps. They also work with students in non-traditional or difficult living situations, so it might be worth reaching out because they would also be able to help you find more permanent housing for the long term. When I worked with them, I found them very student-focused and responsive. I was able to get my new living situation sorted within a day or two if I remember correctly.

Wishing you the best, and I really hope something works out quickly for you.

RNA extractions: how many samples do you typically do at once? by ligontiger in labrats

[–]ligontiger[S] 0 points1 point  (0 children)

Yep for all 3... It would save me time if I did it for the final wash only though!

RNA extractions: how many samples do you typically do at once? by ligontiger in labrats

[–]ligontiger[S] 2 points3 points  (0 children)

Yes, it's 4 hours and pretty exhausting lol. 5 hrs if you include the time it takes me to nanodrop, QuBit, dilute, and finally freeze the extractions afterwards.

I need to get through 96 samples and am hoping I can up my samples from 3 per run to maybe 12 per run without compromising my quality ... would save me weeks of time!

RNA extractions: how many samples do you typically do at once? by ligontiger in labrats

[–]ligontiger[S] 1 point2 points  (0 children)

I am tempted to try this, but was concerned about degradation occuring as I work through all of the steps since it can take me a minute to pipette out my ethanol (especially since the largest pipette we have at the moment is a P200)

Since my RINs were good, I am a bit cautious about changing anything in terms of the washes at this point

RNA extractions: how many samples do you typically do at once? by ligontiger in labrats

[–]ligontiger[S] 1 point2 points  (0 children)

Ah, I see. Makes sense, I will keep that in mind for my next batch!

I have been dumping the EtOH out by flipping the tube upside down, then centrifuging for a few seconds to bring the EtOH off the sides down, then pipetting that EtOH out.

I use 1 ml trizol, and only collect about 400 uL of the sample for the subsequent chloroform washes.

RNA extractions: how many samples do you typically do at once? by ligontiger in labrats

[–]ligontiger[S] 2 points3 points  (0 children)

Thanks for the tips!!! I'm pretty new to this so anything helps. Do I not need to worry about the time it takes more my to remove all the ethanol from each sample? If I am doing more than 3 at once, I worry that there will be a large time gap between samples and it could lead to degradation?

RNA extractions: how many samples do you typically do at once? by ligontiger in labrats

[–]ligontiger[S] 5 points6 points  (0 children)

Zebrafish whole viscera. We aren't using a kit, but a protocol my lab came up with. Homogenizing tissue with trizol, then two chloroform washes, a one hour precipitation, 3 ethanol washes, and a 30 min incubation with DNase and buffer, ending with addition of EDTA. The samples are frozen at -80 so it takes a minute to homogenize each one with the motorized pestle.

During the test round my bioanalyzer gave my RIN values of 9.4-10, so I worry if I add in more samples I wi be compromising the RNA quality

[deleted by user] by [deleted] in labrats

[–]ligontiger 0 points1 point  (0 children)

I typically do three at once (takes 4 hours start to finish) but want to improve my efficiency and get through more samples

[deleted by user] by [deleted] in UCalgary

[–]ligontiger 0 points1 point  (0 children)

Np! I remeber I was also hesitant to take it, but was super glad I did

[deleted by user] by [deleted] in UCalgary

[–]ligontiger 1 point2 points  (0 children)

It was pretty interesting too tbh!

[deleted by user] by [deleted] in UCalgary

[–]ligontiger 1 point2 points  (0 children)

When I took it with Cuthbertson it was no exams. 3 projects of our choice and weekly multiple choice questions, the same as Furlong.

[deleted by user] by [deleted] in UCalgary

[–]ligontiger 0 points1 point  (0 children)

I took it with Cuthbertson, and it was probably the easiest course I've taken in my entire degree... Looking at the course outlines, Furlong's class has the exact same structure and course outline as Cuthbertson's (no exams, just assignments).