hi,
i tried to transfect Tzm.bl to stably express a protein. When i used G418 as selection marker the non-transfected cells didn't die. Anyone knows if for establishing the tz.bl from Hela neo resistance has been used? thank you.
Dear colleagues, I am currently teaching myself in multilevel analysis using educational data. I found few to zero literature on basic analysis methods. In my context we frequently use level-1...
17 February 2021 9,378 5 View
Is there any questionnaire/survey that measures the causes of medical errors? Example: physician negligence vs. systemic issues. Looking for a reliable and validated instrument. Thank you!
15 February 2021 2,874 4 View
I have 2 concentration time curves of a subcut drug implant. the first group has a peak around the 50 day mark where as the second group does not. Both groups received the same implant by the same...
10 February 2021 2,305 3 View
I am currently trying to do image based cytometry from fluorescence microscopy images. From the images I extract data such as cell size and a number of fluorescence intensities. Is there any...
09 February 2021 6,278 1 View
24 January 2021 329 2 View
Hello! I am currently developing an automated counting approach using Fiji to count Collembola from pitfall trapped samples that have been stored in Ethanol (99%) since 2010. I use flotation (tap...
13 January 2021 659 2 View
Hello, I am planning to stain retinal tissue sections for IHC and want to co-stain cones and rods. For this we plan on using a Lectin PNA From Arachis hypogaea (peanut), FITC Conjugate for cones...
12 January 2021 583 1 View
Hi I have a problem where I am trying to vary the stiffness spatially across a structured interface. The whole idea is to vary the stiffness vertically, with a bulk stiffness of 8GPa decreasing...
06 January 2021 262 3 View
Hi, i'm desesperate. i'm trying to produce lentivirus usinf BEI resources kit NR-52948. 24 hours after transfection i detect a massive GFP expression in my 293T cells. After 72 hours i filtered...
30 December 2020 9,889 6 View
Suggestions and recommendations are welcome. A protocol used and experience would also be helpful.
25 December 2020 9,807 4 View
03 March 2021 8,272 1 View
Hello, I would like to ask how to grow E.coli which resists 50 or 100ug/ml of Blasticidin. I have tried many methods by using low salt Luria-Bertain(LB) agar plate. However, no colonies grow. Let...
01 March 2021 2,027 2 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 growing the cells on coverslips and transfecting the fluorescent tagged protein directly on them. Then i want to observe these cells under confocal microscope. Currently i am just using...
01 March 2021 6,142 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
I diluted siRNA and RNAiMAX in opti-MEM and added to the cells which they were in the growth medium. Is it a right way? or should I culture cells in the opti-MEM medium for a while and not in...
26 February 2021 10,041 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 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, I'm looking for a pen/marker to write on the smooth, transparent (non-frosted) part of microscopy slides which does not get washed away by alcohols and most importantly Roti-Histol. I want to...
24 February 2021 9,902 5 View
I tried stable transfection in the U87 cell line with the sgRNA-Cas9 All-in-One lentivector. Cells were cultured in a 6-well plate, 4ug plasmid was transfected after 24hs when the cell confluence...
22 February 2021 2,864 4 View