I want to ask about normalization step: if I want to normalize by DNA content how I should isolate DNA after treatment of the islets with acidified ethanol?
I am staining pancreas with HE. I found beta cells show pale hematoxylin staining in conditional knockout mice compared to surrounding other islet cells, exocrine and acinar tissue " this means...
03 April 2018 363 1 View
Recently, I am trying to isolate islets from db/db mice. I use the same protocol I regularly use for islets isolation from mice but I can't get any islets or very few pale islets attached to...
01 February 2017 5,966 7 View
I am recently extracting murine islets. I see most researchers recommend pooling islets from several mice. I just want to check expression of my protein of interest in islets as an initial stage...
10 November 2016 9,415 3 View
I have been using tubulin as internal control for my western blotting analysis of mice islet lysates. I wonder if there is a better internal control the researchers working with mice islets use.
09 October 2016 6,299 3 View
If this works what kind of tranfection reagent is recommended for the islets?
07 August 2016 4,533 9 View
I have a mouse model that expresses modified Cre recombinase, so that only upon tamoxifen treatment of the animals they express Cre, thus KO of floxed gene can be induced. I want to isolate the...
07 August 2016 2,715 2 View
I am interested in knowing the expression of a certain protein in the islets under feeding and fasting conditions. I use the same total protein concentration for immunolotting. In my first trial I...
07 August 2016 4,351 1 View
I did a glucose stimulated insulin secretion assay for MIN6 cells. I measured the released insulin in the Kreb's Ringer buffer containing certain concentrations of glucose. In the end I collected...
06 July 2016 3,929 2 View
I want to transfect PGL3 plasmid to MIN6 cells. I tried Trans-it trnsfection reagent but didn't work well.
01 January 1970 5,322 2 View
I have reverse sequences (AB1 format), can I base on reverse DNA sequences to perform nucleotide alignment, convert nucleotides to amino acids and deposit the sequence in GenBank database?
11 August 2024 5,138 1 View
After performing symmetric PCR, PCR purification was performed. Afterwards, asymmetric PCR was performed using the PCR purification product as a template, but no ssDNA band was confirmed in the...
08 August 2024 1,668 3 View
I'm trying to find a DNA extraction method for fungi that does not require equipment and heating. Is there anyone who can suggest an alternative option? Thank you
08 August 2024 4,733 2 View
I am currently working on LncRNA; to know the lncRNA-protein interactions I want to do RNA pull down assay, so I need to design primers with T7 promoter. I need assistance in this regard.
07 August 2024 6,622 1 View
Hi all! My thesis groupmates and I are working on purifying an E. coli bacteriophage. The phage was handed down to us by the previous thesis group, but they only managed to purify it to about...
06 August 2024 5,801 4 View
I want to introduce a point mutation (change in one nucleotide) into my gene of interest (DNA binding domain) I have designed primers as recommended on the Data sheet of the kit : -Both primers...
05 August 2024 9,059 3 View
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05 August 2024 4,407 0 View
I have been attempting to extract DNA from Bacterial, Fungal and Yeast banked samples (>1e7 cells) using Prepman Ultra reagent and I seem to be struggling to obtain a sequence. Although the...
01 August 2024 2,079 0 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
99% pure ethanol was used for maceration also alternate methods along with the temperature and time to concentrate the extract can be specified
31 July 2024 5,113 4 View