Single primer PCR - any positive experiences ?

More Bohuš Kubala's questions See All

ATG in pPICZalfphaA vector: Yes or Not in front of the gene of interest ... ?

Is it appropriate to place the ATG codon in front of the gene of interest, since there is a secretion signal that has its own ATG in front of this gene? I need my protein of interest to be...

10 July 2024 3,050 5 View

Is it possible to get gene from Boletus edulis which is more similar to Suillus sp. ?

Hi, I isolated genomes from different fungi. One of them is Boletus edulis (according ITS and 28srRNA primers ~99%). But sequencing of peroxidase gene, which was amplified by specific primers for...

18 June 2023 8,810 0 View

What SUPEROXID DISMUTASE primers do you use for identification of fungal species ?

Hi, I would like to ask what primers for superoxid dismutase do you use or do you know for identification of fungal species ? I used ITS and NL primers but I need to use more markers. I would like...

24 May 2023 2,570 2 View

What light is appropriate for in vitro plant cultures ?

Hi, I would like to ask what light do you recommend to use for in vitro plant cultures? I use full-spectrum LED light (it is pink shining). Is this appropriate for purpose of regeneration of plant...

13 February 2023 6,559 2 View

Fermentation affect activity of Reverse transcriptase?

Hi, I would like to ask, why my reverse transcriptase from fermenter is less active than reverse transcriptase produced from flasks? Thank you for responses Bohuš

24 April 2022 8,738 3 View

Low reverse transcriptase activity from fermenter?

Hi, is it possible, that fermentation Can cause low activity of produced Reverse transcriptase? My RT from Erlenmayer flask is more active. But RT from fermenter Was expressed from T5 system and...

18 April 2022 8,380 0 View

What characteristics belong to Micella formation of anionic alkylbenzene sulfonic acid calcium salt in different solvent?

Regarding the pesticide industry it is a very common and general application of calcium dodecylbenzene sulfonate’s (CaDDBS) usage in the composition of emulsifiable concentrates The calcium...

09 August 2020 3,276 1 View

Cultivation of mycorrhizal fungi on beech roots.

Hi, I'd like to ask about cultivation mycorrhizal fungi which are special beech roots. What media should I use? How to grow in vitro beech ? Thanks for advices. Bohuš

01 January 1970 309 0 View

Reverse transcription Hand-made buffer

Hi guys, Is there anybody who mix Hand-made Reverse-transcription buffers? Is it possible to use Mn ions instead of Mg for better activity? I would be very grateful for All advices. Thank you so...

01 January 1970 7,962 0 View

How much methanol should be added to the fermenter?

How much methanol should be added to the fermenter in which we cultivate Pichia pastoris for the purpose of heterologous enzyme production? We will add methanol continuously with the pump x...

01 January 1970 7,311 3 View

Can I base on reverse DNA sequences to perform alignment, convert to amino acids and GenBank submission?

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 5,136 1 View

How to confirm the site-directed mutagenesis result without performing NGS?

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

I can't see the ssDNA band after performing asymmetric PCR. Is there any way to do this?

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 1,666 3 View

Does crude extraction using NaOH and Tris work well with Fungi?

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 4,731 2 View

Research Methodology - Impact of Corporate Reputation on Stakeholders Behaviors?

Please can anyone support with the survey questions based on RQ measures and propose how to do it in FMCG industry and include as well the role of brand equity Thanks

06 August 2024 946 0 View

Why is my ASO degraded quickly on agarose gel?

I have been working on Red blood cell-derived extracellular vesicles as Antisense Oligonucleotide (ASO) carriers. We normally run agarose gel to quantify the loading efficiency. I used naked ASO...

06 August 2024 3,129 2 View

How much total RNA concentration to be extracted from sorted plasma cells from bone marrow of C57BL/6 mice for RT-PCR ?

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

Anyone having idea about VN primer for miRNA primer design ?

How to design VN primer to attach with universal reverse primer

05 August 2024 2,113 3 View

Why after performing site directed mutagenesis ,I don't see any colony after transformation?

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 9,056 3 View

"A Markov-like Model for Patient Progression"?

A Markov-like Model for Patient Progression" Markov Chain Monte Carlo (MCMC) Markov Chain Monte Carlo (MCMC) is a powerful computational technique used to draw samples from a probability...

05 August 2024 10,079 0 View

Paul Rutland

you could look at this protocol for finding unknown sequence adjacent to a known sequence

http://rothlab.ucdavis.edu/protocols/semirandom.shtml

Junbo Yang

I observed the presence of single-primer amplicons with ONT direct sequencing. However, it is possible that this phenomenon could be attributed to the limitations of the sequencing platform rather than the PCR method.

Atena Abedi

Hi , in selection of mismatches (SNPs) it easily works. Coupling flourcent dyes to such primers can convert PCR to RT PCR .