What are the best ways to reduce generation of chimeric molecules during PCR?

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 9,547 5 View

What is the average disance from a mismatched base for a base substitution occurring during mismatch repair?

Especially in mammalian cells.

05 June 2016 1,312 1 View

Would you like to share with me about isolation of single cells?

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 412 1 View

What percent of template can be synthesized during PCR?

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 4,344 13 View

Does anyone have experience about the efficiency for linear DNA synthesis?

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 5,867 0 View

Which human genes are strongly suppressed by wild-type p53 in human cells?

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 9,495 2 View

Which PCR product clean-up kit you would like to recommend?

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 2,965 7 View

How likely DNA double strands are seperated (melted) at lower than melting temperature?

For example, how many percent (if any) of DNA is melted at 5 degree lower than the melting temperature? Thanks.

11 December 2014 578 8 View

Do you know of any high-yield genomic DNA extraction kit/protocol?

11 December 2014 945 3 View

Is there a way to reduce the interference from genomic DNA on your template-primer pairing?

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 2,314 2 View

Enhanced Yellow Fluorescent Protein (Aequorea victoria) DNA sequence?

I am on the lookout for the Enhanced Yellow Fluorescent Protein (Aequorea victoria) DNA sequence. Does anyone know where I can find it? Thank you in advance

03 March 2021 3,568 1 View

Does anyone have a recommendation for yeast reporter gene plasmid/protocol?

Hi, could anyone recommend a plasmid and/or protocol for reporter gene assay in S. cerevisiae? I want to assess the effect of growth conditions on a transcription factor, so I want to clone it`s...

01 March 2021 210 1 View

Does anyone know HRM channel in a 2 plex QIAGEN Rotor- Gene Q qPCR can be used as a separate channel?

I am going to have 3 different probes in my qPCR work that I am going to do. But I realized that the machine we have in the lab is a Rotor-Gene Q 2plex HRM Platform, saying it has green, yellow,...

01 March 2021 8,544 1 View

Which cell line should we use to perform biological dosimetry experiments?

We are preparing some experiments based on irradiating cells under different conditions in order to evaluate the effects in terms of DNA damage, genetic expression, etc. As our project is...

01 March 2021 3,355 3 View

Is it possible for two human neuronal cell lines to have different gene sequence for the same protein?

I am looking at the ATP1A2 (Sodium/Potassium ATPase alpha subunit 2) in two human neuronal cell lines. Expression levels of this protein seems to be almost equal when detected by one antibody....

01 March 2021 3,607 3 View

Any specific pipeline or parameter to analyse total transcriptomics study of brain tissue samples?

I have two groups of brain samples, control and treated for example. It was total RNA nova seq sequencing. I tried all the available pipeline like: star+rsem+deseq2, Hista+stringtie+cuffdiff,...

27 February 2021 356 6 View

Runs of nucleotides in Sanger sequencing?

I performed site directed mutagenesis, transformation, and then I sent out plasmids for Sanger sequencing and found out that there is extension of DNA just before the stop codon. I am not sure...

27 February 2021 547 3 View

Is there anyone who experienced with cytotoxic effect of Hoescht 33342 dye?

Is Hoechst toxic to cells? In general, it is known as non-toxic. I am working with dsRED MDA-MB-231 cell line. I applied it for my PDMS-based microfluidic platform, for this reason, I incubated...

26 February 2021 9,119 3 View

Calculating volume of DNA required from DNA concentration?

Hi I am a bit confused. They are asking me to find out the volume of DNA required in ul (a total of 30-100 ng for genomic DNA) from the DNA concentration in the nanodrop reading which was 404.8...

26 February 2021 5,029 2 View

Has anyone used the Illumina ITS1 primer sets?

Hi everyone, Illumina provides a list of primers to amplify with high taxonomic coverage the ITS1 region for further fungal sequencing, but I cannot find the exact amount necessary of each...

25 February 2021 6,969 3 View