29 August 2017 6 6K Report

It seems impossible to find on the internet, so far no luck.

More Tomáš Hluska's questions See All
Anybody with experience with whole plasmid sequencing? Should I worry about plasmid oligomers?

I've sent my plasmids for whole plasmid sequencing using the Oxford nanopore sequencing method. All of them were assembled as monomers, but in the reads (see attached pictures), there are even...

19 June 2024 1,280 5 View

What can be yellow colonies after E. coli TOP10 trnasformation?

Last week, I transformed home-prepared E. coli TOP10 cells. When checking the colonies, I noticed that some of them have yellow color. They did not grow on the randomly, but dependently on the...

16 June 2024 3,272 3 View

How to reduce the number of measurements/iterations needed in deep reinforcement learning?

I'm trying to use reinforcement learning with live EEG measurements. However, just 2000 measurements/iterations take 16.6 minutes to measure and it seems I need at least 10 hours of live...

12 June 2024 2,884 3 View

What can be wrong with this agarose gel?

A friend did PCR and then loaded it on agarose gel. All samples were prepared in the same way; the PCR was identical, with the same template, only primers differed, the same amount loaded on the...

04 June 2024 3,049 3 View

How can I see all points in a 5-95 percentile Boxplot in Graphpad?

Hi! I am trying to do some boxplots in graphpad. I would like to use the 5-95 percentile boxplot option, but if I choose this option I do not know how can I also see the individual values of each...

17 April 2024 6,093 0 View

Why could amplicon sizes appear bigger than they should on agaose gel electrophoresis?

I did PCR of 6 amplicons and when I did agarose gel electrophoresis, they all appear consistently bigger than they should be (e.g. the amplicon should be 4000 bp, but is around band of 5000 bp...

09 April 2024 4,487 5 View

What kind of diatom is this?

I was looking at a sediment sample from a "biosphere" that I´m creating and I came accross this little guy. I´m almost certain that it´s a freshwater diatom but would like to know the specific...

19 March 2024 1,763 10 View

Does somebody have experience with ThermoFisher's Platinum SuperFi II DNA Polymerase?

I just found the Platinum SuperFi II DNA Polymerase, which should simplify PCR protocol as it allows uniform annealing temperature of 60°C, but it should also have very high fidelity; >300×...

17 March 2024 7,686 4 View

Me puede proporcionar iso 9001:2015?

quisiera que me proporciones un sistema de gestion de calidad

15 March 2024 3,491 1 View

Iso 9001:2015?

ME PUEDE PROPORCIONAR INFORMACION SOBRESistema de Gestión de Calidad Basado en la Norma ISO 9001:2015 para el Instituto de Educación Superior Tecnológico

15 March 2024 447 0 View

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Illustra™ MicroSpin™ G-25 columns what it is used for?

Hello, We found three packages of Illustra™ MicroSpin™ G-25 columns in the cabinet of an unused lab. They are very old but have never been opened. I have never used this kit before, and I couldn't...

25 July 2024 4,927 3 View

Pink bacterial colonies?

I am cloning an overexpression plasmid with my protein of interest tagged with mScarlet. After transforming my ligated product into DH5α bacteria and plating on LB agar, I noticed colonies with a...

22 July 2024 5,953 6 View

When you express a protein, why do we express not only the domain we want, but also the protein around it?

I want to express STK4, and I've searched the paper for reference. When I check the protein kinase domain sequence for that kinase on Uniprot, it's 30-281, but the paper expresses the protein...

20 July 2024 4,951 1 View

Can I detect the FLAG sequence of my AAV vector by qPCR?

I am treating human cells with an AAV vector encoding for my gene of interest tagged by FLAG. I want to quantify the mRNA expression of my gene of interest via qPCR to discriminate the endogenous...

14 July 2024 8,482 3 View

Unexpected 35 kDa Band Observed During Expression and Purification of Dengue 3 NS5 RdRp Domain ?

Background: I am developing an RdRp assay for Dengue virus-3 and need to express and isolate the NS5 protein's RdRp domain (~101 kDa). My expression vector includes a GST tag followed by a 6x His...

13 July 2024 9,950 3 View

Stable cell line generation: GOI is detected by qPCR but protein is not expressed?

Hello, I'm generating stable cell lines by transfecting plasmid DNA with lipofectamine 3000. The cells went through antibiotics selection for 3 weeks (polyclonal selection) and when I analyzed...

11 July 2024 4,812 5 View

I have constucted a plasmid vector from puc19 with no antibiotic and lacz gene. The problem is screening for transformed cells. Any suggestions?

Get too many colonies, which are mostly non-transformants when tested by colony PCR

11 July 2024 6,775 3 View

Is it possible to generate a (semi-)permanent "transcription bubble" within a replicable plasmid?

I was wondering if it is possible to form a permanent open "ssDNA bubble" similar to a transcription bubble (>13 nucleotides) within E. coli. These criteria are important: 1. Open ssDNA...

07 July 2024 8,275 5 View

What is the Best Midiprep Plasmid Isolation Kit?

Hello, I want to order Midiprep plasmid isolation kit, but I can't decide which brand to use. Can you recommend the brands you always use and think are highly effective? How about GeneJet? Thank You.

30 June 2024 5,865 2 View

Can anyone send me the detailed protocol of lentiviral titer determination and its invitro transduction into Hela cell line or MSCs?

I have searched a lot of information from the published article but couldn't find valuable. As I have prepared the Lentiviral vector and stored it in -80 degree!

21 June 2024 3,484 1 View

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