Iñigo Lobregat

11 Questions 8 Answers 0 Followers

Questions related from Iñigo Lobregat

Can anyone help me with the quality of my gDNA PCR?

Hi, I am trying a PCR reaction that is giving me a bit of trouble. My template is genomic DNA from yeast (s.cerevisiae) and at this point I want to check the quality of the gDNA preps. I have...

02 February 2016 3,710 3 View

Acid wash of coverslips?

When doing acid wash, at some point in the process coverslips are left in HCl 1 M overnight. Yet like this just ONE side of the coverslip is in contact with the acid environment. Therefore how...

01 January 2016 9,725 0 View

Conserved PTM sites?

I want to identify if certain phosphorylation sites are conserved for protein X across humans and yeast. I know from MS data that there are 4 phospho sites in Human protein X. In order to identify...

12 December 2015 2,465 4 View

Any advice on optimizing electroporation?

Hey, I am doing electroporation in Hela cells, but somehow most of my cells die, and my transfection efficiencies are not good. It is the first time i do it thus i wonder if it may be "because of...

11 November 2015 7,935 6 View

Would cut the "post stop codon" sequence in my genes affect the transcription ending and mRNA stability?

I am producing a yeast line with an allele mutated. The protocol to follow will be this one: Obtain gDNA --> clone the desired gene to a plasmid --> quickchange mutagenesis --> introduce...

10 October 2015 9,198 2 View

Is this primer safe?

Hey, I am designing some primers for a cloning. It turns out that I want to use a hexaglycine linker in order to produce fusion proteins, and in order to do so I have to use a primer that would...

08 August 2015 9,149 7 View

How do I design a fusion protein linker?

Hi, I am designing a cloning to produce 2 different fusion proteins. The maps would be: 1- Gene1(1kb)--GeneX(2kb) 2- Gene2(7kb)--GeneX(2kb) My question basically is, which are the principles I...

08 August 2015 2,970 3 View

Is linear DNA stable at room temperature?

Hi, I have left restriction digested and rSAP treated DNA (originally plasmid of 7Kb, now linear of same length) at room temperature for a week. Will it be alright to use, or rather degraded? (or...

07 July 2015 4,456 4 View

How can I keep Hela cells without undergoing mitosis for 24h?

Double Thymidine block is used to synchronize cells. With this system it is possible to obtain cells arrested in the G1 / S phase that will follow cell cycle in a synchronized fashion for more or...

05 May 2015 3,391 4 View

How much lipofectamine plasmid is required for lentivirus production?

I am using lipofectamine to transfect plasmids to produce lentiviruses in 293T cells. The vectors I am using belong to the second generation of lentivectors, pMD2.G (envelope plasmid) and psPAX2...

05 May 2015 7,119 2 View

Anyone familiar with single plasmid detection?

After plasmid transfection is there any detection method that could be used to detect the trasnfected plasmids at sinlge particle resolution? or at least which detection method would be the most...

05 May 2015 7,070 4 View