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Questions related to Cloning
I am trying to ligate a 3Kb band isolated insert to a 12.2 kb vector and I am having issues with transformations. I hardly see colonies on the plates and even if they are formed it is very less....
15 December 2023 9,409 0 View
Hello, I have conducted a bacteria challenge test on tilapia, but I found that I got LD50 negative during calculation. what it means?
14 December 2023 7,023 5 View
Hello, is anyone doing cell culture for shrimp hemocytes? I have a problem with cell sorting; I cannot get the cell sorted out. I hypothesize that I used the L15 medium and did not adjust the...
14 December 2023 6,268 1 View
I am currently conducting research on RSV, and I am eager to know if the number of freeze-thaw cycles affects the titer after amplifying the RSV virus using HEp2 cells. Multiple freeze-thaw cycles...
13 December 2023 915 1 View
Hi, I am currently using Bio-Rad Automated Droplet generator. I am happy with the result. I get on average 19K to 21K droplets per reaction. However, I have also encountered 1 to 2 samples...
13 December 2023 9,186 3 View
for educational purpose.
12 December 2023 4,809 5 View
Suppose I want to prepare 0.1M sodium citrate buffer of pH 3.0, I observed that most people use citric acid monohydrate (as weak acid) and sodium citrate dihydrate (as sodium salt of weak acid)...
12 December 2023 8,360 3 View
Hello dear scientists How can we distinguish maternal contamination from triploidy in QF-PCR analysis?
10 December 2023 9,603 2 View
How to go about testing (in-vitro & in-vivo) the anti-cancer or anti-proliferative activity of a natural plant extract compound against bone/osseous cancer. And which bone cancer cell lines...
08 December 2023 2,315 2 View
I am trying to clone ~3.8kb insert into a ~4.7kb plasmid vector. Things are fine upto transformation. However, during screening when I am isolating the plasmid, I am getting only one...
08 December 2023 6,475 3 View
The liver sections look fragmented, how could the histological technique be improved to better observe this tissue under the microscope?
07 December 2023 7,525 3 View
I am seeking clarification on the differences between the two commercial baculovirus systems for insect expression: Bac-to-Bac system (Invitrogen)and the MultiBac system (from by Imre Berger and...
07 December 2023 2,701 1 View
what is the reason behind this?is it due to the cDNA containation? usually how much cycles we are considering for doing PCR ?
07 December 2023 1,258 0 View
Is there anything can be done if the insert gene contains an additional restriction site which recognized by restriction enzyme not only one restriction site at the end of gene sequence, but...
06 December 2023 8,953 3 View
Hello!! Can anyone tell what could be the reason to get completely opposite Cts value in 2 different qPCR with same qPCR components? I was testing a certain primer on different tissues, first...
04 December 2023 677 3 View
Please tell such properties in terms of the genetic expression of those markers of HepG2 cell lines which makes them a suitable in-vitro model for mitochondrial toxicity & cytotoxicity studies
02 December 2023 4,875 1 View
What cause negative LD50 value and at which conditions
30 November 2023 8,290 0 View
Hi, please i need a help. Attached is my Agarose gel containing ligation reaction (pET28a) and VEGFA (insert). After ligating for overnight at 16 degree and run gel with 10uL from the ligation...
29 November 2023 3,598 3 View
Hello to all researchers, i need to do lentivirus titration and for this I need 10 fold serial dilution i.e 10^-1 to 10^-4...and i have my lentivirus stock stored with me...second thing is if i...
29 November 2023 10,007 1 View
I have run these westerns a billion times and something or the other doesn’t work out. For eg, my gel ran straight and fine but when I transfer the gel to my membrane, half of the membrane shows...
28 November 2023 1,600 0 View
Good day. An immunocytochemical stain for CK20 of urothelium was perfored. Counterstain was Mayer`s hematoxylin (left figure). But after 1 day stain of the nucleouli dissapear (right figure). Why...
27 November 2023 1,378 2 View
My protein is UFM1 tagged with Twin-Strep tag. I double-checked and my tag is not cleaved off or hidden. I made purification with IBA kit (Tactin XT 4Flow high capacity spin kit) with subsequent...
27 November 2023 4,174 3 View
Hello, I want to figure out can I still use my strep-tactin sepharose resin(IBA) if they are dried out for 3 days?
27 November 2023 3,831 1 View
We are working on the expression of resistance genes in potatoes using the LightCycler ® 480. There are two ways to evaluation of the results: basic (that use ΔΔCT method) and advanced (that use...
27 November 2023 8,381 8 View