Wildtype One

fluorescent friday >>> casual friday

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💡 Which Should You Use?

Start with lipofection for common lines.

Switch to electroporation if efficiency tanks or you’re working with tough cells.

Still stuck? Consider viral delivery

(8/9)

🚫 Drawbacks:

• Cell viability takes a hit
• Setup is less convenient (requires cuvettes, buffers, optimization)

(7/9)

✅ Best for:

• Hard-to-transfect cells
• Suspension cells
• Non-dividing or primary cells

(6/9)

2/ Electroporation (physical):

A jolt of electricity makes membrane pores, letting your payload enter directly.

...

(5/9)

🚫 Drawbacks:

• Fails in stubborn cells (e.g., primary cells, T cells, neurons)
• Some end up stuck in endosomes, never reaching the nucleus

(4/9)

✅ Best for:

• HEK-293, HeLa, and other “easy” cell lines
• High efficiency, low toxicity
• Fast, hands-off protocol

(3/9)

1/ Lipofection (chemical):

Cationic lipids or polymers wrap your DNA/RNA and sneak it into cells (usually via endocytosis).

When to use it?

...

(2/9)

Which nucleic acid delivery method wins?

There are 2 major non-viral ways to get DNA, mRNA, or siRNA into cells:

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Just harvested your cells before the flow cytometry run? Avoid common failures with this quick list 👇

this isn’t a test, it’s a mic drop

using cfRNA and microbial RNA signatures for ultra-early cancer detection could redefine how we screen and intervene.

thanks for sharing @sailorrooscout.bsky.social

reminder: small improvements matter when baseline is 1%

nice thread. thanks for sharing @oligogirl.bsky.social

dear santa, my throughput is low and my wrists hurt 🥹

not far off—western blots indeed feel mythically cruel

they worry all of us.

mass spec stocks right now ↗️📈

proof that science does have a soft side

cool stuff @eddpasquier.bsky.social

LCN2 out here multitasking like a stressed postdoc

thanks for sharing @sheencr.bsky.social

finally, maps that actually tell you where you are.

thanks for the major contribution @martinowk.bsky.social

The 8 Western Blot Failures and How to Prevent Them (2025 Guide) Western blotting is a defeating experiment.

this is brutal - we feel you 💔

give this 5-minute article a quick read to save yourself western blot headaches: www.linkedin.com/pulse/8-west...

vibrio just pulled a chromosome infinity stone! 💎

great example of how long-read sequencing still rewrites textbooks @bjesseshapiro.bsky.social

🧫 Join 400+ elite researchers getting weekly lab hacks with our newsletter (it’s free) 👉 wildtypeone.substack.com/about

5️⃣ Important note: In all cases, make sure you report which post hoc test you used—so you don’t leave readers and reviewers wondering!

…and only use a t-test when you have 2 parametric conditions.

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4️⃣ Use FDR (False Discovery Rate) if you have dozens of comparisons. (E.g., gene expression across conditions.)

On Prism, you can find FDR methods (Benjamini–Hochberg and two variants) in the multiple comparisons dialog.

(7/9)

3️⃣ Use Dunn’s test if your conditions are non-parametric (Kruskal–Wallis will replace the ANOVA).

On Prism:

“Analyze” → ANOVA → Choose pairing → “Multiple Comparisons” → “Compare the mean of each column to the mean of every other column.”

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2️⃣ Use Dunnett’s test when you want to compare each group only to the control (not every pair).

On Prism:

“Analyze” → ANOVA → Choose pairing → “Yes. Use parametric test.” → “Multiple Comparisons” → “Compare the mean of each column to the mean of a control column.”

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On Prism: Click “Analyze” → ANOVA (One-way) → Choose pairing → Click “Yes. Use parametric test.” → Click on the “Multiple Comparisons” → Click “Compare the mean of each column to the mean of every other column.”

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Don’t do t-tests—because multiple t-tests = multiple errors.

Do the following Post hoc tests instead:

1️⃣ Use Tukey’s HSD test when you want to compare every group with every other group (all possible comparisons).

Here's where to find it on Prism:

(3/9)

When you have multiple conditions (more than 2), you begin with ANOVA

If your ANOVA is significant, then at least one of the groups is different from the others. But which one?

This is where most people make a mistake...

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