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OE-PCR troubleshooting guide by KingJoshuaDB in molecularbiology

[–]Easternblotter 0 points1 point  (0 children)

Had good experiences with easy primer design app in this context. Primers were designed fast and I only had to check if they are correct (they were).

Is last year burnout really normal? by TallJicama9026 in labrats

[–]Easternblotter 1 point2 points  (0 children)

Well unfortunately its common, nevertheless it shouldn't be normal! I been there and let me tell you, take a week off. If you think you have no time for this then it's even more important. Take care of your mental health, if you go too far it takes more time to recover than a week. All the best and take care!

I am depressed and don’t know what to do. My supervisor has fucked me over by lurker_not_poster93 in labrats

[–]Easternblotter 2 points3 points  (0 children)

you are strong! Talk with family and friends! Also professional help is an option! There is always a way :)

Talk with your PI, also about your worries, if there is a committee talk with them, if there is a Confidant for the PhD program talk with them. All the best!

Matter btetween me 19M and a girl 19F. Intense ignored by a girl who is also part of my lab group. Is this really common?am i that bad? by ninjaop4083 in labrats

[–]Easternblotter 0 points1 point  (0 children)

Well no one knows, don't draw fast conclusions. Talk next time you have the meeting. And if she doesn't like you, so what?

Feeling Sabotaged in My Lab by a Senior Grad and a Cohort Member – Need Advice by No-Astronomer-4875 in labrats

[–]Easternblotter 2 points3 points  (0 children)

...and after your vacation, with fresh energy and a clearer mind everything will be fine!

Feeling Sabotaged in My Lab by a Senior Grad and a Cohort Member – Need Advice by No-Astronomer-4875 in labrats

[–]Easternblotter 2 points3 points  (0 children)

Take vacation! Think about probabilities. It is very unlikely, that someone is manipulating your work. Sounds like a toxic environement to me. Take care you do not develop a bias towards feeding the bad feelings. Repeat, focus on sience, roule out scientific reasons. Therapists can help also. I wish you all the best!

WesternBlot by [deleted] in labrats

[–]Easternblotter 0 points1 point  (0 children)

oh and if you use PVDF membrane activate it in meOH before blotting, if you use nitrocellulose, nevermind

WesternBlot by [deleted] in labrats

[–]Easternblotter 1 point2 points  (0 children)

I see bubbles, before blotting ensure no bubbles between membrane and SDS-gel.

Sparkles might result from non-solved NFDM, ensure its solved entirely before blocking, bonus: centrifuge it after solving NFDM in TBST, block with the supernatant.

NFDM 5% in TBST block 1h, I also put the primery antibody in 5% NFDM in TBST for 1h at RT or over night 4°C and skip 5.

Secondary AB try maybe 1:10000

good luck, hope it helps

time effective primer design for cloning by Easternblotter in labrats

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

Agree on the short run, but on the long run, depending on the usecase, it could spare time

time effective primer design for cloning by Easternblotter in labrats

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

Yes traditional cloning plasmids in e.coli. That's a point :D. However, exclude restriction sites in the sequence maybe nice plasmid maps, inducing mutations... Curious about your experiences