Magdolna Keszthelyi

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Questions related from Magdolna Keszthelyi

NEBuilder HiFi DNA Assembly Mix usage, directly on PCR product: does it really work?

Hello Everyone, the description of NEBuilder HiFi DNA Assembly mix says: "No PCR clean-up step required". I always used this mix after DNA extraction from agarose gel , but now we have some...

22 June 2023 6,250 1 View

Total RNA isolation, cDNA synthesis - is it possible, that there are just short fragments of cDNA, and it is impossible to get longer PCR product?

Total RNA was isolated from C. elegans, and cDNA was reverse transcribed. We can get nice PCR product whenever we amplify a housekeeping gene, or a specific one, but only short ones (maximum...

20 January 2022 7,447 4 View

What type of microinjectors are available in Europe, that can be applied for C. elegans injection?

We are very beginners in this field, so far we have transformed the worms with Gene gun. Now we would like to buy a microinjector equipment with all accessories, that are needed... I can not find...

15 December 2020 5,753 0 View

Can C. elegans MosSCI system be used with gene gun (microparticle bombardment), or it is exclusively compatible with injection?

We would like to generate a single copy insertion of a human gene in C. elegans genome, and we decided to try the MosSCI method. The problem is that we have a gene gun (BioRad PDS-1000/He...

28 June 2019 9,420 0 View

Why does Myfi polimerase make errors in a 2-3 kb long PCR?

Hello. We are trying to clone a2-3 kb long insert into a 3,5 kb long vector by NEBuilder. We had to amplificate by PCR both the insert and the vector. We used MyFi polimerase.(...

13 March 2018 7,608 4 View

Can anyone help in planning cloning vector primers?

I would like to make an expression vector, and attach a tag to the 5' end, so the start codon will be moved upstream. My forward primer will contain a restriction site, the kozak (with the START),...

01 March 2017 5,440 3 View

Does alpha-tubulin immunostaining show primary cilium in epifluorescence microscope?

I found a lot of fluorescence pictures of primary cilium stained with alpha-tubulin antibody in the literature, but I am not sure if they made by confocal or epifluorescence microscope? Will I see...

17 February 2017 8,346 2 View

How to differentiate HK2 cells?

In the literature I found this: "after 3 day post-confluency, cells are differentiated." But how? With or without FBS? If with, than 1, 5, 10% or changing? How to prevent outgrowing? Thanks

30 January 2017 4,377 3 View

If mRNA level is zero, why we should look at protein level?

I know that mRNA level often do not correlate with protein level. But, if mRNA level is zero, than I think protein can not be anyway. Why we should see it at protein level as well? thanks

09 January 2017 5,651 5 View