15 Questions 6 Answers 0 Followers
Questions related from Bohuš Kubala
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...
11 July 2024 3,050 5 View
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...
19 June 2023 8,810 0 View
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...
25 May 2023 2,571 2 View
Hi, I would like to ask if anybody has positive experiences with single primer PCR ? Can you recommend me any proven protocol of this type of PCR ? Thank you for all recommendations. Bohuš
09 March 2023 4,642 3 View
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...
14 February 2023 6,559 2 View
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
25 April 2022 8,738 3 View
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...
19 April 2022 8,380 0 View
Hello, I'd like to ask whether it is necessary to add primers or oligo(dt) to Reverse transcription mastermix (MMLV RT)? Is it possible to mix Primer independent RT reaction? If you have any...
01 January 1970 6,605 2 View
Can Taq DNA polymerase uses RNA as its substrate? I mixed RT reaction with: dNTPs, RNA of GFP gene, Reverse transcriptase and water and incubated 30 min at 45 °C. Then I mixed PCR with Taq DNA...
01 January 1970 7,558 4 View
Hi, I did heterologous expression of reverse transcriptase in different strains of E. coli at 3 temperatures, 37 °C, 28 °C and 20 °C. As I expected, BL21 provided the best solubility at 20 °C and...
01 January 1970 2,422 1 View
Hi, I have problem to sterilize freshwater plants (Anubias, Bucephalandra). I used chlorine salts and ethanol. I got contaminations. I think AgNO3 could works. Do you have any other protocols for...
01 January 1970 2,807 5 View
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
01 January 1970 10,001 0 View
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 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 7,312 3 View
