Iis there any shuttle vector for yeast and E.coli teransformation?

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Has anyone had luck using cell lines other than HEK 293 (such as COS1) to package lentivirus?

I am looking to package lentivirus in endothelial cells but I am opposed to using 293 cells due to personal ethics regarding the source. I have done some reading and saw that COS1, COS7, & CHO...

25 February 2021 3,030 1 View

What is a good reagent for lentiviral plasmid transfection in HeLa cells?

I have been using Lipofectamine 2000 but was hoping there may be a better protocol or one for something like Lipofectamine PLUS reagent.

24 February 2021 1,633 4 View

Hi, Good day! Do you have related studies about my title, "Factors Affecting the Senior High School Students' Stress Tolerance in Problem Solving"?

I am new researcher. I want to learn more and I need your help. Thank you.

04 February 2021 6,834 4 View

Efficiently transfecting HeLa Cells?

I am currently looking to transfect HeLa cells using lentivirus and Lipofectamine 2000 and have been looking for cited protocols. Would anyone have any that they are willing to share?

31 October 2020 5,639 2 View

How to compute the effects in half fractions?

Greetings, I'm doing a screening design and want to allow for lack of fit. I need replications so I want to add four additional points to my dataset. Rather than use center runs I use corner...

11 October 2020 9,672 1 View

Which of the ff. solvent is more effective to use as a bactericidal agent in antimicrobial testing of staphylococcus species, ethanol or methanol?

The antimicrobial activity testing of staphylococcus species.

02 October 2020 6,996 7 View

How to get pure recombinant virus with plaque purification ?

We are working on getting pure desired gene transfected virus. We used YFP fluorescences tags to distinguish the virus with or without gene replaced. Using PCR testing, we always have wild type...

01 August 2020 2,087 1 View

What types of research methodologies involve working with or observing people, but do not require IRB approval?

I am interested in understanding more about NGOs that work with refugees. I am brainstorming a project that does not involve IRB approval.

31 July 2020 1,854 5 View

How do you remove your name you did not do for a conference presentation?

I got an email from someone I do not know saying my name is on her conference presentation. How do I remove it? We have the same first initial and last name (although now she has a different last...

12 January 2020 2,877 4 View

Multiple energy minimization steps in gromacs?

Hi all, I have a rather naive Molecular dynamics question, though functionally useful. I am simulating a rather large structure on a congested HPC, and was wondering if there is an accepted...

30 November 2019 9,877 2 View

Did anyone ever try using pLVX-TetOne Plasmid with a copGFP as a selection marker instead of Puromycin?

When using a lentiviral vector for inducible expression of genes using the Tet-On system. In the literature, I saw a widely used of pLVX-TetOne-Puro Plasmid, is there a reason why people prefer to...

25 February 2021 1,011 3 View

How to use protein A G beads to purify antibody?

Secretory universal influenza antibody CR9114 were successfully expressed by adenovirus vector ,then we used protein A G beads purify CR9114.But the molecular weight of the purified antibody under...

22 February 2021 4,318 2 View

Covid 19 Pandemic ' impact on digital transformation?

Covid 19 Pandemic and digital transformation

22 February 2021 3,351 5 View

What are the best cations for preparing competent cells, Ca2+ or Mg2+?

I have been reading various articles and most state that treatment with CaCl2 produces higher efficiency of transformation than MgCl2. Can anyone explain why Ca2+ is better than Mg2+ at...

16 February 2021 8,519 5 View

Is the logarithmic transformation a necessary step in time series analysis ?

Hello, I'm checking if the log-tranformation of time series data is a necessary step; We all know the aim of such transformation, but in every study (in time series analysis) we found the authors...

15 February 2021 5,655 14 View

How much amount of DNA (ligation mix) should I use for transformation?

Hi Fellow Scientists! I am trying to clone my 300 bp insert into a 6 kb vector. I run a few different ligation reactions using (1:3 vector: insert molar ratios) 10, 20,30, and 40 ng/ul vector...

15 February 2021 9,563 6 View

How to produce huge amount of protoplasts from Fusarium graminearum?

Hi, all I am using some protocol for protoplast production from Fusarium graminearum (Fg), but I need them in a huge amount. Sometimes the protocol worked well but most of the time failed....

14 February 2021 2,286 4 View

How to measure 50uL of cells?

Hello, I am planning to do a transformation, and on the protocol it says "Scrape a 50 ml volume of yeast from the YPAD plate using a sterile inoculating loop or sterile toothpick". But I can...

14 February 2021 6,247 3 View

Mixed ANOVA with some normal and some non-normal data?

I am looking to run a 2x5 Mixed ANOVA to examine Sensory Processing Sensitivity in Highly Sensitive People (HSP) and Non_HSP. My Between-Subjects factor is HSP and Non-HSP. All participants tested...

11 February 2021 1,445 6 View

Any probe or assay for the detection of Cysteine in yeast invivo ?

Dear fellow researchers, I would like to detect the cysteine levels for yeast in vivo. I would really appreciate if anyone could suggest me any fluorescent probe or assay kit available. Currently...

11 February 2021 3,292 3 View

Karen A. Darbinyan

Hello, see links below may be useful for you.

Good luck!

https://www.sciencedirect.com/topics/neuroscience/shuttle-vector

https://onlinelibrary.wiley.com/doi/full/10.1002/yea.3228

Article IncP Plasmids Are Unusually Effective in Mediating Conjugati...

Ali Javadmanesh

Dear Mary

You have a lot of options to choose a suitable shuttle vector for your purpose. It depends on a lot of factors! Size of your inserts, MCS enzyme, selection marker, induction system and ... . Here you can browse and filter your search parameters which fits your need: https://www.addgene.org/vector-database

Good luck

Navnit Kumar Ramamoorthy

Dear Mary,

I have seen people using the pAUR shuttle vector for yeast-E.coli combinations. It has the Aureobasidin A selection marker in it. The link below may help you

https://www.takarabio.com/products/cloning/linkers-primers-and-cloning-vectors/cloning-vectors/e-coli-yeast-shuttle-vectors

Pamela A Marshall

I'm old school and use these.

Article A System of Shuttle Vectors and Yeast Host Strains Designed ...