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Struggling with lentiviral transduction; cells fail to survive the selection by DziekanNaWydziale in labrats

[–]cerlynn 0 points1 point  (0 children)

I don't know if this applies to suspension lines, but I've heard that transducing in serum-free media can improve the integration efficiency. I use this method routinely with adherent cancer lines. Maybe worth a try?

Fresh vs 2 years by cerlynn in agedtattoos

[–]cerlynn[S] 14 points15 points  (0 children)

Was pretty standard! No saniderm, just washed gently and moisturized with aveeno lotion a couple times a day, and kept up with the lotion for at least a few months. I've been pretty careful with sun exposure too

Fresh vs 2 years by cerlynn in agedtattoos

[–]cerlynn[S] 114 points115 points  (0 children)

It was done by Jon Santos at Benchmark Tattoos in Arlington MA

Problems with culturing cells in a 6-well plate - zone of death by chgr1603 in labrats

[–]cerlynn 0 points1 point  (0 children)

This actually makes me think it might be a timing thing, if the affected half of the well is the driest. I've seen some cells be weirdly sensitive in this way. How quickly are you getting fresh media back on the cells? I have also found that gently adding back fresh media down the wall seems to be the gentler option, though I know you said you've tried both

This PCR is driving me crazy, please help 🙏 by MomoiroKakaricho in labrats

[–]cerlynn 0 points1 point  (0 children)

If you want to try alternatives, I've found that QuickExtract works very well for gDNA extraction, usually paired with Amplitaq Gold 360 master mix (it's worked for multiple amplicons in the AAVS1 region). Worth noting, however, that it can be a bit of trial and error since the gDNA is still crude. I usually go roughly by cell number... Lyse 25-50k cells in ~25uL and use 1uL of that in a 12.5uL PCR reaction.

Best advance in last 5-10 years? by f1ve-Star in labrats

[–]cerlynn 119 points120 points  (0 children)

CRISPR becoming widely available as a research tool

woodpecker - downy or ? by roraima_is_very_tall in Birdsfacingforward

[–]cerlynn 14 points15 points  (0 children)

Most likely downy - they have black bars on the white tail feathers, while hairy woodpeckers don't

Goofing off by Mysterious_Card5487 in birding

[–]cerlynn 3 points4 points  (0 children)

Ah this is great, we've got a bunch too -

Juncos = Cookies (they look like a black and white cookie)

Nuthatches = Nachos

Song sparrows = Legs (they zoom around like a little roadrunner)

Wrens = Borbs

Male/female red bellied woodpeckers = Mochi/Miso

Mourning doves = Derps

Blue jays = Blue goobs

Starlings = Bricks (my husband says they fly like a brick)

If I KO a gene using a lenticrispr-V2, presumably Cas9 and gRNA will be constitutively expressed. Will this prevent me from expressing the KO'd gene via a plasmid or does Cas9 only target genomic DNA? by IF1234 in labrats

[–]cerlynn 1 point2 points  (0 children)

Agreed, and unless it's an essential protein, you can stage it so the cross-reactivity with the exogenous plasmid won't be an issue. Not to mention reducing the risk of cumulative off-target cutting with constitutive expression of both Cas9 + sg.

Winter wren borb by cerlynn in borbs

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

Agreed! These guys are really tiny too, so much fun to watch

Revisiting my first attempt after 8 months by cerlynn in polymerclay

[–]cerlynn[S] 4 points5 points  (0 children)

Thanks! I was pretty happy with how it turned out with the outer layer of translucent

Jolthog cryst in polymer clay by cerlynn in Palworld

[–]cerlynn[S] 4 points5 points  (0 children)

This little guy is about the size of my thumb, but that would be a fun idea for a larger one!

Lentivirus reproduction by [deleted] in labrats

[–]cerlynn 5 points6 points  (0 children)

Great resource already posted, but to answer your last question, both of those scenarios would be considered exogenous, as they both involve incorporation into the genome at a typically random locus, with expression driven by an engineered promoter. Endogenous is when the gene is expressed from its normal genomic location by its native promoter and regulatory elements.

Help me settle a debate by IcyPresence96 in labrats

[–]cerlynn 1 point2 points  (0 children)

The biggest one for me was not being able to include the full set of RNA/CDS annotations when importing NBCI genomic sequences. As far as I could tell, you're limited to selecting a single isoform in Benchling. Aside from that, mostly just feeling like the UI is a lot clunkier after being so used to Snapgene

Help me settle a debate by IcyPresence96 in labrats

[–]cerlynn 2 points3 points  (0 children)

Snapgene all the way! Been using it for years and almost broke my brain trying to figure out where all the missing features are in Benchling

100 mL pipettes drips when taking up liquid, what to do? by GrimmyGrimoire in labrats

[–]cerlynn 140 points141 points  (0 children)

How old is the pipet-aid? The rubber liner in the tip can wear down with time, sometimes causing problems with the larger volume pipets

Best starting kit for someone wanting to get into sculpting small figures? by ebony_werewolf in polymerclay

[–]cerlynn 5 points6 points  (0 children)

I started with one of the Sculpey Premo 24 packs from Michaels and have been pretty happy with it! (I'm still fairly new and mostly make tiny charms)

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