I ordered Arabidopsis mutants from ABRC and found that one of the mutant is hetrozygous (confirmed by genotyping), later when I performed qPCR, I did not see any amplification of that particular gene (undetermined values). What can be the reason ?
I want to know whether my over-expression line is homo or heterozygous in genetic background. I know some methods but want to is there any way to find that. Methods I know- 1. Check segregation...
07 August 2018 2,012 0 View
Hello, I am looking for a protocol by which I can detect the Ion-leakage in the anthers of my Arabidopsis Mutants. I want to know the detailed protocol mentioning the tissue amount (anther...
05 June 2018 1,590 5 View
By mistake I kept some for my flower bud samples in paraformaldehyde for more than a week (usual time if few hours), can i still wash them with 1x PBS and store them in 70% ethanol at 4 C...
10 November 2017 4,280 1 View
I am performing PCR for my several isolates, but in one isolate I'm getting multiple bands after the amplification. What could the reason be? I'm using a single set of primers.
09 October 2013 787 36 View
I'm cloning a fragment of 3200 nts into plasmid. The cloning was successful, however, 02 amino acids were mutated. Now I want to fix these 02 aa by site-directed mutagenesis technique using...
08 August 2024 4,645 2 View
I've been performing RNA extraction on cotton petiole tissue for a few months now using the method described in the following paper, a derivative of the typical hot borate method...
08 August 2024 9,882 2 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 have sorted anti-NP specific plasma cells from bone marrow of C57BL/6 mice at certain times after immunization with variable counts and isolated total RNA using TRIZOL method for RT-PCR using...
05 August 2024 8,835 1 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
Hello everyone, I performed a PCR yesterday, and the results showed no bands on the gel. Of course, I probably missed some crucial steps, like adding my samples to the PCR strips themselves, for...
31 July 2024 2,406 6 View
Can anyone with an MJ research / BioRad PCR machine from ~2010 or earlier tell me the external measurements (LxWxH) of the removable standard PCR alpha modules that can be removed from a PTC200 or...
30 July 2024 2,867 0 View
I have been following Rivero et al 2014 protocols, but I am not sure when I should stop watering because my plants are completely yellow but the siliques are not 80% brown. Any recommendations?...
30 July 2024 2,301 7 View
Hello all, I have been trying to follow a 2-stage PCR protocol used to amplify barcodes of a large yeast library, as per Nyugen et al. (2022) -...
30 July 2024 841 2 View
First, thanks for taking the time to read my puzzle 🥹! I am currently working on the methodology part of my dissertation, and I have chosen a questionnaire as the instrument for my quantitative...
27 July 2024 3,457 3 View