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

Have you applied 3D Laser scanning and Building Information Modeling for facility management during operation phase?

14 March 2020 7,806 3 View

Is there any difference between Authoritarian Mother and Authoritarian Father?

Good day! I'm looking for journal articles that would help me differentiate between an authoritarian mother and an authoritarian father. Thank you in advance!

28 February 2018 5,212 3 View

Cost-effectiveness model: Converting different rates on different rates into one probability?

Hello Everyone, Need quick help! I have 5 different rates for recurrence from 5 studies with different followup. How do I combine them to find one probability to be used in cost-effectiveness...

05 November 2017 4,107 7 View

Are there known antibodies produced when a human is infected with STH (Ascaris lumbricoides, Trichuris trichiura, hookworm)?

Are there known antibodies produced when humans are infected with STH (Ascaris lumbricoides, Trichuris trichiura, hookworm) and do the produced antibodies differ per helminth? Thanks in advance!

19 September 2017 1,116 5 View

How long we can keep soil water extraction in refrigerator (4oC) ?

07 August 2017 6,149 6 View

Is there an assay that is able to quantify the degradative enzymatic activity of fungi without isolating the enzyme from the fungi?

18 May 2017 8,694 3 View

What are the implications of neutral theory for interpreting patterns of genetic variation in ecology and evolution?

04 November 2016 7,331 4 View

Any changes in soil dissolved organic matter if we conduct second time extraction by hot/cold water?

06 July 2016 8,229 3 View

Any changes in soil dissolved organic matter if we conduct second time extraction by hot/cold water?

05 July 2016 2,288 3 View

Why does blocking with my immunogenic peptide cause nuclear fluorescence in ICC?

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

Does Napabucasin soluble in dichloromethane and in ethanol?

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

Help request in CST microwave?

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

Issues with Permeabilization during Flow Cytometry?

Hello Everyone. Currently I am working to characterize macrophages in the myocardium after ischemia-reperfusion injury in rats. Due to the low total cell number isolated from rat hearts I can...

01 March 2021 3,867 3 View

Is CD44 degraded in lysosome together with hyaluronan after CD44 mediated endocytosis of hyaluronic acid?

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

Dapi fluorescence protocol for colon cell lines ?

28 February 2021 2,534 2 View

Does RNAiMAX transfection reagent work in serum free medium?

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

Has anyone had luck using cell lines other than HEK 293 (such as COS1) to package lentivirus?

I am looking to package lentivirus in endothelial cells but I am opposed to using 293 cells due to personal ethics regarding the source. I have done some reading and saw that COS1, COS7, & CHO...

25 February 2021 3,030 1 View

Unknown debris sheets observed in cell culture flasks?

Hi, We culture neural progenitor cells in T175 culture flasks coated with PLO-Laminin. We have successfully grown these cells with our protocol for months. Recently, we observed large sheets...

25 February 2021 3,443 4 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