How can I repress an endogenous gene expression?

I want to down-regulate the expression of a gene in yeast. Could you please suggest me some methods? Thank you.

11 December 2018 620 0 View

Which enzymes catalyze NMP, NDP and NTP transforamtion?

I want to know the mechanism of NMP, NDP and NTP transformation in eukaryote. Can anyone suggest me some papers? Thank you!

05 June 2017 4,440 0 View

What is the mechanism of yeast integrative plasmid?

05 June 2016 4,813 5 View

How can I condon optimize a protein?

I want to get a K. lactis yeast condon optimized Cas9, which software or website can do that?

04 May 2016 6,034 2 View

What is the promoter of Kanamycin in prokaryote plasmid?

I can not find Kana promoter in the map, e.g., pet28, while AmpR promoter is always indicated. Then what is the Kana promoter sequence? And can E. coli containing a plasmid with KanMX(TEF promoter...

03 April 2016 5,452 6 View

How can I add a tag right bebind the 3' end of a gene?

01 February 2016 2,699 2 View

Can anyone suggest me a good cotton genome database?

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02 March 2014 1,888 0 View

Can anyone help with vector construction with PGBKT7?

I want to insert the CDS of my gene into the MCS of PGBKT7, but I found that the vector contains two sites NdeI(CATATG) and NcoI (CCATGG), I would like to know whether the ATG in these two sites...

02 March 2014 2,777 3 View

Are there any "master transcription factors" in yeast?

Are there such TFs in yeast, like Myc, whose overexpression could affect multi-pathway protein expression?

01 January 1970 8,719 1 View

Enhanced Yellow Fluorescent Protein (Aequorea victoria) DNA sequence?

I am on the lookout for the Enhanced Yellow Fluorescent Protein (Aequorea victoria) DNA sequence. Does anyone know where I can find it? Thank you in advance

03 March 2021 3,568 1 View

Does anyone have a recommendation for yeast reporter gene plasmid/protocol?

Hi, could anyone recommend a plasmid and/or protocol for reporter gene assay in S. cerevisiae? I want to assess the effect of growth conditions on a transcription factor, so I want to clone it`s...

01 March 2021 210 1 View

Why might my GFP tag be dissociating from my plasmid after transfection?

I have a set of stably transfected cell lines all transfected with plasmids containing GFP tags on the C terminus. During a western blot using anti-GFP antibody, one of my plasmids has dissociated...

01 March 2021 9,310 4 View

Expression cloning by using cDNA,do I need to modify the cDNA first?

I am going to have a expression cloning of mammalian gene by using shuttle plasmid to transforming the E.coli However I don't know I should only inserting the Coding sequence ,or I can...

28 February 2021 5,440 3 View

Problem with shRNA transfection. Why are only two plates getting contaminated instead of all?

I transfected my LNCaP-WT cells with 3 shRNA plus their NTC two weeks ago and split two puromycin selected cell plates on Friday last week(Feb 26). I checked for GFP in the cells, and they all...

28 February 2021 4,949 3 View

PUC19 as a transfer plasmid for lentiviral delivery?

Hello, Is it possible to use pUC19 as a transfer vector to be packed in using the second generation viral particles packaging system( pMD2.G; psPAX2 plasmids)? As far as I understand it there is...

28 February 2021 4,868 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

Why do the PCR bands from the cDNA templates (extraction from the cell after transfection) show different size from the band of the plasmid template?

After transfection with the plasmid ( linearized ) and subcloning of the cell lines, RNA was extracted from the cell and then reverse transcripted to cDNA. When PCR reactions were run to verify...

25 February 2021 5,712 3 View

How do I transfect a large plasmid into PC12 cells with nerve growth factor (NGF) receptor?

I have a large 16 kb plasmid, which I need to transfect into PC12 cells. Lipofectamin 2000 didn't work and with GFP alone the transfection rate is very low. I also tried the neon invitrogen...

25 February 2021 6,635 5 View

What's the concentration recommend for recombinant protein use in western blotting?

I'm currently working having problem with the concentration for recombinant protein use in western blot, the one been used is Recombinant Human CD19 Fc Chimera Protein-rndsystem which does not...

24 February 2021 8,514 3 View