Subham Basak

10 Questions 32 Answers 0 Followers

Questions related from Subham Basak

CHO-K1 suspension adaptation protocol?

I am trying to adapt adherent CHO-K1 cells in F12K+10%FBS to suspension culture in only F12K, but in vain. I have followed Mirus bio as well as Thermo protocols, please suggest any alternative....

01 August 2024 3,711 0 View

Why cannot i find my protein on cell surface after antibiotic selection of expressing plasmid?

I cannot confirm cell surface protein expression by flow cytometry even after transfection and antibiotic selection of cells. Does it take long for proteins to express on the cell surface? the...

29 July 2024 3,134 3 View

How do we isolate only B cells from Spleen/ lymph nodes for Hybridoma generation?

As we know, lymph nodes have a mixed population of cells, how do we ensure only B cells are getting isolated and subsequently fused with myeloma cells? Thanks in advance!

31 January 2024 3,412 2 View

Why ExpiCHO cells need 8% CO2?

Why do ExpiCHO cells need 8% CO2 levels in the incubator? Is is because they have high metabolic rate, thereby producing more lactate and needing more CO2 to buffer the media? Would like to know...

27 August 2023 6,677 3 View

Wnat is the size range of ExpiCHO cells?

Also would like to know the ExpiCHO parameters like ellipticity/ circularity and max/min. diameter to count the cells via automated cell counters. Thanks for the help!

27 June 2023 881 0 View

Why do dead cells in 6 well plate collect at the centre?

I am doing HEK293 antibiotic selection after transfection, and everyday i find dead cells collected at the center of the well, although the plate is kept static in the incubator. Any plausible...

26 May 2023 4,486 4 View

How can i increase biomass of my E. coli culture to better plasmid yield?

Hi everyone, I want to increase the biomass of my E. coli culture for better plasmid yield. Use of different strains of cells and media or any other suggestions are welcome. Thanks!

12 October 2020 6,554 3 View

Would you recommend using SYBR green based real time PCR kit for disease diagnostics?

It is seen that most diagnostics kits are Taqman based, now as PCR is quite sensitive, can we settle ourselves with SYBR based kits in a resource-crunch set up?

18 August 2019 3,191 3 View

Is the amount of RNA extracted from normal human serum too little, to give amplification of a marker gene by RT PCR?

I have isolated RNA from 50ul serum and eluted the RNA in 30ul of DEPC- treated water. But when i am preparing 10ul of cDNA from 1ul of the RNA and using it for amplification of a 115bp...

23 January 2019 6,942 3 View

COVID-19 Detection

I would like to get learned insights into detection of COVID-19 using techniques beyond the routine RTqPCR from you all. Thanks for your thoughts!

01 January 1970 5,348 1 View