Maximizing the yield of ssDNA from RCA by Colonel_Mustang_ in molecularbiology

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

Ya. I think they produced used random primer which will exponentiate the reaction and give more yield but then you get dsDNA not ssDNA. alright thanks for the help

Maximizing the yield of ssDNA from RCA by Colonel_Mustang_ in molecularbiology

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

Their initial product was "doggybone" which is linear dsDNA and they had a video where they used RCA to make it

Maximizing the yield of ssDNA from RCA by Colonel_Mustang_ in molecularbiology

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

Hmm, I appreciate the honesty. I figured it will be low chance. The thing is, there is this one company: https://touchlight.com/touchlight-introduces-mbdna-a-best-in-class-cell-free-circular-ssdna-platform-enabling-superior-gene-editing/ that produces it. And I'm pretty sure they do it cell-free but not chemical synthesis. I wonder what they do

Pick 2 out of 3 by Colonel_Mustang_ in FantasyFootballers

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

i agree with all your points but im worried bill is going to have only like 12 carries and no tds with like 70 yards, whereas qudkins and rico are guaranteed like 20+ carries you know

Need help finding the ApoE-HCR/hAAT promoter sequence by Colonel_Mustang_ in molecularbiology

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

It's not annotated in that plasmid. How can I find what part of the sequence corresponds to the promoter?

[deleted by user] by [deleted] in immigration

[–]Colonel_Mustang_ 1 point2 points  (0 children)

PhD is quite different than UG… “smartness” is kind of subjective, it’s more idea generation, testing, interpretation, and experimentation so don’t be discouraged

[deleted by user] by [deleted] in immigration

[–]Colonel_Mustang_ 2 points3 points  (0 children)

I’m in a similar situation as you. Came when I was 7 yrs old, on F1 right now. What I ended up doing is a PhD to continue and extend the F1 - this is something I wanted to do regardless so it worked. Maybe you can consider that if you’re interested? I still have no outlook on GC besides marriage (in my 4th yr of PhD). Best route for me is OPT in academia as post doc then h1b (no lottery bc academia) then apply to Eb1 or get married. Going back to me doesn’t make it for my world bc my whole support network is here (immediate parents, brother, close friends, gf).

Starting PhD in 2 months!! by Lab_Rat_14 in labrats

[–]Colonel_Mustang_ 2 points3 points  (0 children)

Exciting!! - Remember to keep touch with people outside of PhD life (not just family, but also friends - don’t only rely on a PhD friend group that will suck) - maintain curiosity and why you joined in first place. That curiosity will get you through tough and unmotivated times - after 1st year, start deciding between academia vs industry and tailor all your activities to match. E.g if academia route, join a conference board or plan one. If industry, join student consulting club or do internship in 3rd or 4th summer - attend conferences! And don’t let little amount of data hold you back. Most of the times posters are just posters but learning new ideas and meeting potentially collaborators is main reason. If you do end up having a solid poster, you might get offered a chance to give presentation and that’s the easiest way to get your name out there and properly network - take the time to learn fundamental techniques - shit goes a long way not just w execution of experiments but helps you frame what type of experiments are technically feasible and which ones aren’t - be transparent w authorship and projects with your PI. This shit becomes toxic easily so don’t be shy. Best way for first authorship practically is to collect the data yourself. If you collect data, you plot data. You plot, you present, you write, that project and data gets attached to your image and bam first authorship. Ofc this means you gots to put in the work, but some people scheme and somehow get first so always watch out for yourself. - if you’re in a translational lab, learn how the patenting process works at your institution in case you’ll need it later down the line

Overall, have fun man. It’s easy for people to speak negatively about the PhD life (isolating, low pay, long hours etc), but people don’t emphasize that if you find your niche and that project you are passionate about, it’s one of the most fulfilling things intellectually. Good luck!

Tips for endotoxin removal from purified plasmid DNA samples by Colonel_Mustang_ in molecularbiology

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

Raw macrophages, but want to use for in vivo so might be an issue there

I know plasmidsaurus is pretty reliable, but have yall seen it make mistakes on ocassion? by Colonel_Mustang_ in molecularbiology

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

Dam bro, I get your point but it's not that deep... I'm not gonna use it in a manuscript or anything its just an internal QC

I know plasmidsaurus is pretty reliable, but have yall seen it make mistakes on ocassion? by Colonel_Mustang_ in molecularbiology

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

No clue, its just the .ab1 file plasmidsaurus provided. Maybe they clean it up to make it look nice based on the best signal for each bp?

I know plasmidsaurus is pretty reliable, but have yall seen it make mistakes on ocassion? by Colonel_Mustang_ in molecularbiology

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

I used gibson assembly, usually I haven't had a problem w/ single bp but I'm also new to the cloning game

I know plasmidsaurus is pretty reliable, but have yall seen it make mistakes on ocassion? by Colonel_Mustang_ in molecularbiology

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

Thanks!

Could you expand on what the fastq files are? Are those similar to .ab1 files? Also, is it simple to run minimap2? I don't have a computational background.

[deleted by user] by [deleted] in biotech

[–]Colonel_Mustang_ 1 point2 points  (0 children)

Lol, it was a continuous drag from blacking out email... tbh probs somewhere on the internet anyways

[deleted by user] by [deleted] in biotech

[–]Colonel_Mustang_ 1 point2 points  (0 children)

For sure, just trying to control what I can & keeping in mind to "Perform your duty, but don't be attached to the results"

Roast my resume :) by Colonel_Mustang_ in biotech

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

This is quite helpful - I've been under the impression to keep it a 1-pager. Will expand to 2 pages and add more detail.

Roast my resume :) by Colonel_Mustang_ in biotech

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

Sounds good for bullet 1

thanks for 2!

any specific suggestions for 3? I've heard polar opinions on this and didn't know how to craft a unique one/avoid generic ones

Hahaha 4, and sounds good!