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

Agarose gel purification pre MinION sequencing?

Dear all, we performed an agarose gel purification with MinElute kit (QIAGEN) to extract bands between 3500 bp and 6000 bp for further sequencing with MinION. After the purification process,...

16 February 2021 5,178 1 View

How can I concentrate peptides?

I am working with an initial volume of 20 mL* and using a 10 kDa filter to separate proteins from peptides. With this filter I am able to concentrate the proteins upstream of the membrane....

15 January 2020 1,520 7 View

Is it possible to freeze whole but with RLT buffer for future RNA extraction without RNA quality loss?

Hi all! I was wondering if anyone has experience or information about storing whole blood samples in RLT buffer for future RNA extraction. In my case, I will need to process/store a lot of blood...

09 October 2019 7,925 7 View

Why is one plasmid not transfecting HEK 293t cells , while a simliar second plasmid sucessfully transfects the cells with the same protocol?

I ordered and produced a plasmid from Addgene containing PIK3CA and GFP (pHAGE-PIK3CA-C420R (item #116457). I performed mini- and midi preparations and verified plamid production with a...

02 October 2019 4,078 2 View

Does anybody know the name of this plant from the Canary Islands?

15 April 2019 1,244 6 View

Any data regarding Calcium levels in glioma cells cytoplasm after adenovirus infection?

We are performing experiments on glioma cells. After infection of a cell line (U251), we see that Calcium levels increas even using a mock virus. Any reference that could explain this? Maybe...

03 March 2019 7,772 1 View

Input of WRF wind ouput to a CFD?

Hi. I am trying to a mesoscale and microscale simulation using WRF and a CFD. The output from WRF will be used as the boundary input. I was wondering though since WRF's output for wind is by grid....

21 January 2019 4,990 2 View

How to express units of DPPH radical scavenging activity?

Hi! we are doing some measurements about the DPPH radical scavenging activity of seaweed extracts. We know that the main units used to express this activity are in %. But we have different sample...

31 December 2018 1,744 7 View

How to calculate change in temperature on realistic resistor ?

When using the Specific Heat Capacity ,the following equation provides the temperature change: (T2-T1) = IIRt /(mc) I-Current V-Voltage R-Electrical Resistance t-Time m-Mass of the material c-...

12 December 2018 5,329 2 View

Can linear DNA be used as template for site-directed mutagenesis?

Is it possible to induce site-directed substitution mutation by quick-change method on linear dsDNA? or it has to be cloned in some vector? If yes, should it be treated with the Dpn1 enzyme...

03 March 2021 401 4 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

Expression cloning by using cDNA,do I need to modify the cDNA first?

I am going to have a expression cloning of mammalian gene by using shuttle plasmid to transforming the E.coli However I don't know I should only inserting the Coding sequence ,or I can...

28 February 2021 5,440 3 View

Difficulty in cloning of 1kb gene into vector?

I am facing difficulties in cloning a 1kb gene into a vector (pJIT163). I have my gene on interest (GOI) in pUC57 and want to clone in pJIT163 using SalI and BamHI restriction sites. I am getting...

25 February 2021 3,221 7 View

In what ways do fellow reaerchers think that mRNA vaccination may be under CRISPR systems technology application ?

There are many ways for mRNA to intervene with genome expression. This question is raised for experts in CRISPR systems. Any RNA does not stand alone within acell. There are many ways to become...

18 February 2021 9,109 5 View

Why are Sanger Sequencing reads interrupted in some edited clones?

I have a Crispr/Cas9 edited bulk population and I can briefly check the indel ratios in the bulk population using Sanger sequencing. After cloning single cells, I wanted to check the indel types...

18 February 2021 8,198 6 View

Hi, can anybody help me with the suggestions how can I amplify a stress induced gene of 1107 bp?

I am trying to amplify the TGA1 (Transcription factor 1 ) of Arabidopsis ,which is 1107 bp. I have treated the plants with Salycylic solution over night and extracted the RNA and make cDNA. I...

17 February 2021 7,802 2 View

Alternatives to traditional cloning of insert into backbone (dsDNA)?

Hi, I am stuck with a cloning experiment: The insert & backbone(bb) share one compatible end at 3' end(NotI) and one incompatible end at 5' end (XbaI for insert; HindIII for backbone). So i...

16 February 2021 1,664 3 View

Why do I have trouble with cloning duplicate gene in the same plasmid?

Hello! I am trying to generate a construct in this format: seqA - seqB - linker - seqC - seqB into pSF-CMV-Ub-Puro-Ascl plasmid 0.5kb 2kb 66 0.25kb 2kb I have tried many...

15 February 2021 7,118 3 View

Would using an asRNA for repressing transcription of a high expression Listeria monocytogenes non-coding RNA harbored in a plasmid be a viable option?

Hello all, I am interested in deleting a highly expressed non-coding RNA located on a Listeria monocytogenes plasmid. Currently, there are not any optimized gene deletion methods for Listeria...

09 February 2021 4,994 1 View