Sarmishta Majumdar

9 Questions 25 Answers 0 Followers

Questions related from Sarmishta Majumdar

How to carry out a kill curve experiment?

Could someone please help me understand how to carry out a kill curve experiment? This is for antibiotics G418 and Puromycin for CHO cells

06 June 2023 2,305 0 View

Does AvrII and XhoI produce complementary overhangs?

Hi, I have a vector that has AvrII (at 5' end) and XhoI(at 3' end) RE sites at the cloning site for my insert. I wish to know if these two enzymes give rise to overhangs that are complementary or...

01 March 2023 8,527 5 View

How to check if the ATP is hydrolyzed in the ligation buffer? Does the ATP form any white ppt at the bottom of the tube, and dissolves after mixing?

Since my ligation is not working (no colonies in insert: vector plate) and analyzing my results where we could find that my digestion is just fine and same in case of my transformation (lawn in...

20 January 2023 2,633 8 View

Can double digested vector and insert be stored and used later for ligation, after a few weeks?

I have my vector and insert double digests purified and gel extracted. After ligating and transforming, I didn't observe any colonies on the plate. Is it okay if I use the purified double digests...

05 January 2023 8,219 3 View

Ligation not working, reason?

I have tried to ligate my vector+insert in ratios of 1:3 and 1:5, along with keeping a vector only control. Although, I got colonies post transformation the no. of colonies is more or less same in...

02 January 2023 6,218 21 View

Antibiotic range to be tested in Kill curve for CHO-S cells?

The antibiotics are Neomycin, Puromycin, Zeocin, Blasticidin

31 July 2022 2,353 2 View

Puromycin kill curve for CHO-S (suspension) cells

Could anyone suggest a protocol to determine the conc. of puromycin required to kill CHO-S cells? If anyone has performed earlier, can you please suggest a range of puromycin to be tested?

01 January 1970 2,008 6 View

Suggestion for collaborations in my PhD research: Vector optimization and Cell line engineering

Hi, I would like to approach Professors/ researchers from my field i.e. mammalian cell line development and vector optimization/ molecular biology to guide me in my project. I have around 4 years...

01 January 1970 1,338 2 View

Fed-batch study using freestyle CHO cells producing a recombinant antibody

Has anyone performed a fed batch study using freestyle CHO-s cells in shake flask? I've few pools of recombinant antibody producing pools derived from freestyle CHO-S and I wanted to understand...

01 January 1970 1,292 1 View