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Questions related from Tany Nino
Hi, I am learning to perform footprinting experiment. In the protocol I have there are the following components of the reaction buffer: HEPES buffer (7.8 pH), glycerol, BSA, DTT, KCl, MgCl2,...
29 May 2024 1,254 0 View
I'm having difficulty finding a possible solution to my problem: I have a plasmid encoding a protein. In the nucleotide sequence of this protein, I need to add a nucleotide sequence (highlighted...
26 May 2024 6,807 3 View
I know that to many of you this might seem like a really stupid question, but I am a beginner with PCR. If I have a DNA sequence (for example, a plasmid) into which I want to insert a small DNA...
24 May 2024 3,638 1 View
Probably this question seems fool to you, but I am quite new in the research area and there are a lot of experiments I have never done. My question regards the amount of buffer, enzyme, incubation...
16 May 2024 3,963 1 View
I attempted to clone a small DNA insert (~100 bp after digestion) into a plasmid of unknown size, which originally carried an insert of 111 bp that I removed by double digesting the plasmid. After...
15 February 2024 2,742 3 View
Hello everyone, I am currently working on expressing a protein in Rosetta 2. After harvesting and freezing the cells, the next step is to purify the sample. The initial step involves resuspending...
16 January 2024 9,807 3 View
I inoculated 20 mL of Rosetta pre-culture into 1L LB broth supplemented with 1 mL of Kanamycin and 1 mL of Chloramphenicol. After one hour of incubation, I checked the OD, and the value observed...
14 December 2023 4,893 1 View
Dear all, I used both mass photometry and SEC-MALS to define the mass of my recombinant protein. Based on the amino acid sequence only, it should be 92 kDa. However, it holds different glycan...
28 July 2023 4,303 2 View
Dear all, I am troubled with my CD data (it is just my first experience with it). 1. How do you interpret my result? I can see just a negative signal at 208 in the above graph (alpha helices?). 2....
28 June 2023 1,941 0 View
Hi, I hope to receive your help. I am a student, so I do not have a lot of experience in research. If you wanted to understand how RNAP, ribosome, and transcription factors work altogether and...
28 May 2023 8,151 0 View
I am using some restriction enzymes, and according to the protocol, the amount of enzyme should not exceed 10% of the total reaction volume to avoid star activity. This activity is due to the high...
24 May 2023 4,106 2 View
I am using mammalian cell cultures to express a protein. Which concentration of Vit C should I use to add in the culture to "help" my mammalian cells?
10 March 2023 6,826 1 View
