I am currently optimizing electroporation conditions for mouse embryonic stem cells. What should be ideal range of pulse duration?
Attached there are some parameters you can set for electroporation: as you will see it depends on the type of cells and cuvette and temperature. You have to find the ones that best fit your system. Good Luck
This method seems to be easy, sensitive and cost effective. All I am looking for internal control DNA. Also, please suggest any other easily available PCR methods or commercial kits
01 February 2019 9,048 2 View
I have transfected mouse ES cells with single gRNA, Cas9 system. I am able to detect cleaved bands after T7 endonuclease. However, how to determine if this is monoallelic or biallelic mutation?
11 December 2017 878 4 View
I wish to know electroporation conditions especially for transfecting Cas9 RNA and protein
11 December 2016 2,484 2 View
I have tried amplifying my individual fragments using Phusion but it is not working consistently but always works fantastically with KAPA. I think I am unnecessarily wasting time on Phusion....
05 June 2016 9,302 4 View
I have tried doing a PCR with kapa as well as Phusion HF polymerase. I am getting amplification with kapa, but Phusion is just not working. Both the master mixes use same: 1.5mM MgCl2 and 0.2mM...
04 May 2016 6,915 3 View
Hi all, I am validating a polyclonal antibody raised against human ANT4, through WB and ICC. In most of my controls, the antibody shows very little off-target binding, but when I pre-incubate the...
03 March 2021 5,999 3 View
I need to prepare a solution of Napabucasin (inhibitor of cancer cell stemness) in dichloromethane. The drug is expensive, so maybe somebody already tried to do it. If it is soluble, what is the...
02 March 2021 9,945 3 View
Question to you and THEM, the New Journal, "Integrative Psychological and Behavioral Science" -- do you not know, and have you not seen, this done before? There appears to be a core problem for...
02 March 2021 3,024 2 View
im working now on TPV solar cells and metamaterials and i realy need our help in CST microwave i have a tpv cell simulation and i want to extract particullarly the absorbtion of only the active...
02 March 2021 2,246 2 View
Hello, The Cranet website does not provide any lead to the data obtained from their multinational survey on HRM practices. Any help on accessing the data would be most appreciated. Thank...
01 March 2021 9,536 3 View
As the organizations getting prepared for the fourth industrial revolution, to enhance the skills of their workforce what type of changes are necessary and going to occur in the training and...
01 March 2021 3,199 3 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
Also when RHAMM binds hyaluronic acid, they get internalized, will RHAMM also be degraded? Or both CD44 and RHAMM will be transferred back to the cell membrane? Asking for breast cancer cell line...
01 March 2021 8,169 2 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