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

Can plasmid used for S.cerevisiae transformation be used for Kluyveromyces lactis?

03 April 2016 2,497 5 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

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

Precipitate after adding laemmli buffer to protein solution containing Potassium Acetate?

Hi, I am running a size exclusion chromatography experiment with a buffer containing Potassium Acetate as a salt. I analyse these fractions through SDS-PAGE. After boiling my SEC fractions in...

01 March 2021 2,622 3 View

What is the most suitable server for predicting protein structures in bacteria?

I have used the i-Tasser several times, however it has been unavailable for several days. I tried the swiss-model, but the output was not very pleasant due to the model used. Is there any other...

28 February 2021 4,521 3 View

Do you need the CMV enhancer before the CMV promotor for a proper function of CMV on the gene that follows?

I'm a student and I have to produce a cell line with a knockin for the NRF2 gene with GFP. I have to put a promotor in front of the GFP because the gene will be too far away from the promotor of...

28 February 2021 7,127 2 View

Are protein samples in SDS compatible with ELISA?

I have used an AllPrep DNA/RNA/Protein Mini Kit QIAGEN kit to extract RNA and protein from my samples. At the end of the protein purification, I resuspend my protein in 5% SDS. Will these samples...

28 February 2021 7,370 3 View

Any specific pipeline or parameter to analyse total transcriptomics study of brain tissue samples?

I have two groups of brain samples, control and treated for example. It was total RNA nova seq sequencing. I tried all the available pipeline like: star+rsem+deseq2, Hista+stringtie+cuffdiff,...

27 February 2021 356 6 View

How to replicate the well conditions of a crystal hit with 20%(w/v) isopropanol?

I had a crystal hit in a well with 0.1M HEPEs pH 7.5, 10%(w/v) PEG 4000, and 20%(w/v) isopropanol. Why is isopropanol in w/v here if it's just solvent? How do I recreate this well condition?

25 February 2021 9,761 1 View

Problem with ligating 2.8 kb insert into 8.5 kb plasmid?

I am trying to clone 2 copies of the same mammalian gene into the pSF-CMV-Ub-Puro Ascl (contains HindIII, KpnI and NheI cut sites) plasmid. This is what the final product is supposed to look like...

24 February 2021 6,310 3 View

What urea buffer can i use for biotagged protein extraction from streptavidin beads?

Hi all, I have been doing research on biotagged LDB1 proteins in Mel cells. I purify the proteins using M280 streptavidin beads. The yield is very low so I've been trying to optimize it by...

24 February 2021 5,749 3 View