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Hi, I have recently attempted natural competence transformation with Bacillus subtilis (WB800N), following the protocol from Mobitec. I used 5ug/ml chloramphenicol for my selection. My...
25 June 2022 9,146 0 View
Hi, I was experimenting with different amount of seeded RAW 264.7 cells incubated with LPS to find out which seeding density to use for my upcoming experiments. I was using the 10k, 20k, 40k,...
15 June 2022 7,511 0 View
Hi, This will be my first time doing this assay. For context, my experiment will be to determine if some compounds have any anti-inflammatory effect. I plan to do the following steps: 1) Seed...
26 May 2022 576 3 View
Hi, I just started using this strain of B.subtilis, and during transformation (Heat shock, natural competence), I noticed the negative and positive control plates looked exactly the same (I am...
25 February 2022 1,826 3 View
Can you autoclave 100% glycerol straight up to sterilize it? Will it result in caramelization or will it be fine? I am aware I can dissolve it in water and autoclave it but it would make things...
16 February 2022 4,504 2 View
For context, I am doing multiple restriction digestions (>50) and I want to streamline it by making a 10x cutsmart buffer + restriction enzyme mastermix. Reaction volume will be 50ul so the...
18 October 2021 1,139 4 View
I did a sequential digestion of ApaI and BamHI on a few of my plasmids, using 4ug each (plasmid size ranged from 4kb to 9kb), followed by gel extraction. All but 1 of my plasmid ,the biggest one...
20 April 2021 5,005 0 View
SDS PAGE experiment had a weird result. Are there any ideas as to why the gel turned out like this? Could it be that the gel was very dense or the acrylamide mixture was poured unevenly resulting...
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