I mean the toxicity is based on DNA sequence or its structure per se not the transcription or expression products from the DNA.
The only example I know of are plasmids for SF3b1:
http://onlinelibrary.wiley.com/doi/10.1111/j.1742-4658.2011.08387.x/full
There it seemed as the sequence of mouse SF3B1 was toxic and then Yokoi and colleagues made a mouse/Xenopus hybrid which could then be successfully cloned.
The chimeric molecules generated during PCR are really bad for our study on recombination, since the chimeras look exactly like recombination. I wonder what and how (would be better if protocol is...
06 July 2016 9,640 5 View
Especially in mammalian cells.
05 June 2016 1,427 1 View
I am interested in all the methods or instruments used to isolate single cells especially from mammalian tissues. Any experience of you is appreciated.
09 October 2015 501 1 View
For example, the parameters for PCR are denaturing at 95, annealing at 58 and elongation at 72, while the melting temperatures of primers are around 60.
04 May 2015 4,456 13 View
We are trying to synthesize mouse DNA using "linear" synthesis in very limited number of cycles without "denaturing". Do you have any experience for linear DNA synthesis, especially about the the...
11 December 2014 5,960 0 View
For example, how many percent (if any) of DNA is melted at 5 degree lower than the melting temperature? Thanks.
11 December 2014 688 8 View
I am interested in looking for those genes that are expressed more than five fold higher when p53 is lost or mutated.
11 December 2014 9,593 2 View
I am trying to clean the PCR products for deep sequencing, and I would like to know which kit of which company you think is the best (based on your experience if you have done). Thanks.
11 December 2014 3,077 7 View
I am trying to extract genomic DNA from a small pool of mouse cells (around 60,000 cells) and the yield is critical for me (at least 80%). Does anyone have good suggestions for which genomic DNA...
11 December 2014 1,042 3 View
For example, whether long DNA fragments or shorter ones (such as with restriction enzymes) interfere with the pairing of the primers to the templates of interest with different efficiency or not.
11 December 2014 2,415 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
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'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 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
What is the most suitable experimental setting to understand the consequences of a particular gene: (1) targeted degradation of the specific transcription factor (TF) of the gene, followed by...
04 August 2024 6,265 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
I have an RNA-seq data that I have analysed using Limma-voom and have extracted the gene IDs, log2FC and the p-values. At p value < 0.05, I have over 10,000 DEGs, however, when I run the GO...
31 July 2024 225 2 View
I am currently investigating the cytotoxicity of a series of herbal extracts, and like many studies, I have been using the MTT assay to evaluate cell viability. However, I am encountering a...
31 July 2024 193 4 View
I am currently working on a project involving liposomes and need to determine the maximum volume of siRNA that can be added to a 2.5 mL liposome solution with a total lipid concentration of 10...
30 July 2024 6,420 1 View