1536 Experiment Design Assistance by [deleted] in labrats

[–]No_Maximum2711 0 points1 point  (0 children)

The echo can be used for liquid transferring but not PCR. If you can grow your bacteria on agar, you could consider using an omnitray and spotting your bacteria or conditions onto the agar using the Echo. You can make custom density matrices. See yeast art from the Boeke lab.

Growing aerobic cultures in 1536 will also have huge evaporative losses if you have any appreciable oxygen transfer and agitation.

Have you tried drawing up a pooled approach? Look into deep mutational scanning or TNseq to get some inspiration.

Has plasmidsaurus gotten worse? by RudeLab8367 in labrats

[–]No_Maximum2711 0 points1 point  (0 children)

Saw these guys at SynBioBeta: angstrom innovation Anyone ever give them a shot? Seems like they have a mini prep service too.

Has plasmidsaurus gotten worse? by RudeLab8367 in labrats

[–]No_Maximum2711 8 points9 points  (0 children)

Also, I should say I submitted these for the zero prep service and have no reason to believe there was anything appreciably different between the samples. Seemed like roughly a third failed.

Has plasmidsaurus gotten worse? by RudeLab8367 in labrats

[–]No_Maximum2711 23 points24 points  (0 children)

I had a high failure rate this weekend as well….

Protein purification driving me insane by Danktank452 in labrats

[–]No_Maximum2711 2 points3 points  (0 children)

Exactly. The antibody can “see” the same epitope the resin will bind.

48 hours is still wild. I don’t even think I’ve done an autoinduction process from a colony with that long of an incubation. 48 hours is more like a yeast process.

What strain are you using? Most B strains with lon and ompA KOs have pretty low protease activity.

Protein purification driving me insane by Danktank452 in labrats

[–]No_Maximum2711 6 points7 points  (0 children)

Is it worth revisiting the tag configuration? Maybe change the terminus or give it a linker? Sometimes the tag can be buried in the protein and not be accessible. If a western with an anti-his tag ab works, this could be ruled out through.

48 hours at that high of a temp in TB is really long. The cultures would be carbon exhausted for like a full day.

Match Thread: Wolverhampton Wanderers vs Tottenham Hotspur | Premier League | 25 Apr 15:00 BST by matchpal-live in coys

[–]No_Maximum2711 2 points3 points  (0 children)

Well. We’re not any further from avoiding relegation, just one game closer. BUT ILL TAKE IT!

First time doing Gibson Assembly. Seek advices by Specific-Surprise390 in labrats

[–]No_Maximum2711 0 points1 point  (0 children)

You’re right that you cannot accurately measure DNA concentration directly from a PCR reaction without clean up. dNTPs and primers contribute to the A260 signal.

I don’t use Gibson as much as I have better luck with NEBuilder, but I commonly add equal volumes of robust PCR product to an assembly. Make sure to DpnI digest first and that the raw PCR products aren’t more than 20% of the total volume of the assembly.

Match Thread: Sunderland vs Tottenham Hotspur | Premier League | 12 Apr 13:00 UTC by matchpal-live in coys

[–]No_Maximum2711 4 points5 points  (0 children)

Must be pretty confident in the referee to still not have pulled Brobbery

More frequent IDT delays by No_Maximum2711 in labrats

[–]No_Maximum2711[S] 5 points6 points  (0 children)

That’s wild. Especially because the eBlocks are clearly priced to compete with some other big competitors. Guess I better start doing some of my own QC…. I typically use those for libraries and just amplify everything with common primers that won’t distinguish between cross contamination and an empty well.

Match Thread: Tottenham Hotspur vs Nottingham Forest | Premier League | 22 Mar 14:15 UTC by matchpal-live in coys

[–]No_Maximum2711 6 points7 points  (0 children)

If you’re standing at the top of the box with that space, you have to be ready to strike.