As above
I want to use two promoters to express two gRNA in a single plasmid and wish them to have same number(i.e. two promoter with nearly same strength). I am not sure whether I should insert a...
05 June 2019 6,972 1 View
I want to redo a RNA extraction from the E.coli cells and I store the liquid culture (in TB) for two days. I am not sure whether I can still use them or I need to regrow some cells.
01 February 2019 7,854 2 View
I was expecting only ~110bps amplicon in my sample, but I see a very broad band. Attached is the result. 2 and 4 only contain the template, no primers. Also, I think my intensity is kind of low.
10 November 2018 6,768 0 View
I want to compare transcription of a gene both inserted in the genome and a plasmid. Should I extract the RNA in the exponential phase or stationary phase?
10 November 2018 2,019 11 View
I want to count the amount of crRNA expressed in a plasmid with Ecoli. I am not sure whether I can use a RT-qPCR to do it. The length is between 50 - 100bps.
08 September 2018 1,113 6 View
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
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
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
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
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
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
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
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
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
I have been trying to have a good AAV titration for my production and I have tried different plasmids to set up as a standard curve but each one behaves differently. i am trying to use ITR for...
23 February 2021 5,167 3 View
