Hi all, does any of you know of specific primers for both emetic and diarrhaeic forms of Bacillus cereus?
A need to analyse specific sequences gathered from genbank in order to analyse primers matching/designing. I have used Sequencher (gene codes) in the past and I loved it. Yet prices are now...
24 April 2024 259 3 View
Having problems with dimers and an answer from Paul Rutland for another question made me think that the fact that I store my PCR mix in the fridge might be the problem. I must add that in this mix...
28 March 2024 9,886 3 View
I have a dataset of trace elements LA-ICP-MS analyses with raw data, which consists of a set of a single file for each spot analysis. Each of these files has some columns (masses) and plenty of...
24 March 2024 898 6 View
I have so far disliked Taqs that come with dyes because dye concentration seems excessive to me. I have recently bought a Taq with dye-free buffer, I wonder if I can use a small bit of gel loading...
20 March 2024 3,193 0 View
Hi all. I have some primer pairs which always produce those horrible primer dimer bright smear! Increasing annealing temperature does not solve the problem. Any suggestiopn for a PCR enhancer or...
19 March 2024 6,067 4 View
Need do look for milk bacterial pathogens using PCR. Does anyone suggest me a good protocol for DNA extraction? I have heard milk proteins are difficult to deal with aabd act as taq inhibitors
13 March 2024 402 0 View
I am making tests for diagnostic PCRs. At the moment I am trying to standardize a pcr reaction for a circa 180 bp product. I did not get positive bands so far. Interestingly, I get what appears to...
26 February 2024 2,820 3 View
We would like to preserve RNA from mammal blood samples. Unfortunately the only product I am aware is RNA Later and it is very very expensive for our limited budget. Any ideas? Thank you very much
12 December 2023 6,163 2 View
Sorry basic question. I am aware primers should be described in the 5'-3' direction in the literature. When I am ordering primers I have taken from published papers, should I use the same sequence...
18 October 2023 5,523 3 View
We are currently having problems with PCR amplifations of a ~200bp gene fragment. We cannot see any product after a 35 cycles amplification round and usually use a simple reamplification protocol:...
27 September 2023 9,212 1 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
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 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
How to design VN primer to attach with universal reverse primer
05 August 2024 2,116 3 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
It's an end-point PCR protocol. I'm using 1.5% agarose gel with SyBR Safe dye and TBE as a running buffer, visualization on BioRad XR+ system. I was primarily thinking of primer efficiency,...
01 August 2024 4,673 4 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
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