Have you ever seen a small number of cells, 50-100 thousand, after centrifugation as a dry pellet in an Eppendorf tube without a microscope? Are there methods to potentially stain the cells in the suspension so that they can be visible?
The source of classification bias in marker gene metagenome sequencing? a variability in the taxonomy classification of microbial communities when using different primer pairs (e.g. for 16S rDNA)...
29 June 2024 1,048 1 View
How to store SDS-PAGE for fluorescent imaging without fixatives? I am running protein samples on SDS-PAGE that are tagged with fluorophores. If I place the gel in a destain solution of...
29 June 2024 8,720 1 View
Is a 5` phosphate needed for homologous recombination? I am a bit confused whether phosphorylated DNA is needed to achieve efficient homologous recombination in e coli. I believe there are methods...
06 June 2024 7,065 1 View
What is the reason or importance of diluting an enzyme before an experiment? I'm writing a report on enzyme activity and characterisation and i need help with understanding this: we were told to...
06 June 2024 8,714 2 View
So, here is what I am dealing with: The Tm of the primers is 55 for the forward one and 56 for the reverse one. In -silico PCR produces 750 BP band, but in reality I get 3 bands. The third one...
17 March 2024 9,914 5 View
I am amplifying target sequence 450 bp. I get single sharp band in the control and faint in sample with another sharp nonspecific product. why? I need to get one single band from my sample to...
17 March 2024 804 3 View
I am doing a CRISPR targeting to knock out one intron of my gene of interest in a cancer cell line and replace that intron with GFP. I am wondering if I should pick single cell clone with possible...
17 March 2024 9,713 1 View
For example, If a PCR primer can detect ~100 fg of DNA, how we will calculate it to Bacterial copy number?
17 March 2024 1,816 2 View
How can the design and synthesis of guide RNA molecules be optimized for improved CRISPR-Cas9 gene editing?
05 August 2023 1,375 2 View
I could not find a study that used CRISPR-mediated HDR for stable integration of large genes. I would be grateful if anyone could point me in the right direction.
05 August 2023 486 2 View
Hello, I am currently having problems with RNA extraction. I am using mouse liver (C57BL6J), and I have extracted RNA from mouse liver before. Before this experiment, my final RNA pellets were...
11 August 2024 7,082 3 View
I am not able to get good literature and the physics behind how first these grains and grain boundaries arises out of no where when we make a pellet to study its dielectric properties and then how...
07 August 2024 5,177 3 View
I am trying to prepare samples for IP. The pellet size is good and I am using 1% Tx-100 and using a syringe to further lyse the cells and maintain protein-protein interactions. I keep having a...
01 August 2024 7,199 1 View
I'm fairly new to making homemade comp cells and have done so with some mild success. What is generally an acceptable way of resuspending comp cells during wash steps? I've been told to swirl the...
31 July 2024 8,309 0 View
I am cloning an overexpression plasmid with my protein of interest tagged with mScarlet. After transforming my ligated product into DH5α bacteria and plating on LB agar, I noticed colonies with a...
22 July 2024 5,953 6 View
How to determine tannin content in fodder pellets?
22 July 2024 8,163 3 View
I am extracting proteins from plant samples and am currently attempting to use the TCA/Acetone extraction method. I am having trouble resolubilizing my sample after drying the pellet. I collected...
15 July 2024 9,025 3 View
Someone advise me to use 1 to 2mm undersized pellet but it is not available. Do anyone have any experience in the field of tunnel kiln ?
28 June 2024 6,355 0 View
Hi, I´d like to trace the C and N flow in a recirculating aquaculture system by labelling the fish feed with concentrated stable isotopes. From a supplier, I found a product ouf of algae lysate...
19 June 2024 746 0 View
I've pelleted down the overnight ampicillin supplemented cultures of DH5ALPHA for pET22b isolation 5 days ago and stored them on -20°C without adding any buffer or glycerol. Would the cells be...
09 June 2024 6,479 2 View