I designed the primers for qRT PCR, but doing standard curve it shows the primers are not gene specific. I tried NCBI and other soft wares too. any suggestion please.
CRISPR/cas9
10 November 2018 2,050 0 View
I want to find the genes related to development of female insects. I have the transcriptomic data, How I find? which pathways are related to development? Your consideration will be highly appreciated.
06 July 2018 663 0 View
I run the standard curve to know the primers specificity for amplification. I got this curve and melting peak. kindly explain whether it is right?
04 May 2018 8,349 0 View
I run the qRT PCR to detect the target gene expression with different time points. each sample have three technical replicates and compared with control. Now How I calculate the value of cq? how...
03 April 2018 9,223 2 View
I want to observe the larval and pupal stage of moth, for this rear them on artificial diet. Your kind help will be appreciated.
03 April 2018 1,573 2 View
We treated the adult moths and observed the egg hatching which was significantly reduced compared with control but the mating behavior was not changed. If we want to study the mechanism from...
01 January 1970 9,224 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