Ks Lim

10 Questions 25 Answers 0 Followers

Questions related from Ks Lim

What is the maximum deletion size is applicable to achieve in site-directed mutagenesis?

I am try to delete a domain containing 660bp using Takara PrimeSTAR Mutagenesis Basal Kit. However, sequencing results show no success in deletion of this region. Is the size too big for deletion...

13 September 2018 2,283 5 View

The best culture condition for Kit-225

I am currently culturing the T-CLL, Kit-225 cell line. My culture condition is RPMI 10% FCS with 20ng/ml IL2 (R&D, about 420IU/ml). Cells look viable but proliferation rate is slow. Is there...

28 June 2017 2,054 6 View

How can I overcome such problematic melt curve in qPCR?

I am facing problem in qPCR analysis. The file attached shows the melt curve of GAPDH. In my setting, my template was 100ng and primer concentration 500nM in 20ul. Cycling number was 40 cycles. In...

02 January 2016 8,175 4 View

How to achieve better amplification in qPCR?

I am currently comparing ULBP-4 expression in cancer cells. The method I used for qPCR as follow: a) cDNA synthesis = 1ug of RNA in 20ul b) I diluted the cDNA 10X prior to qPCR c) I used 4ul of...

31 October 2015 4,108 8 View

Can I culture unstimulated NK cells without IL2?

I would like to culture natural killer cells harvested from patients PBMC without IL2 for a day. Will that cause massive cell death?

21 March 2015 7,393 7 View

Can formalin mix with Triton-X in PBS to make 3.7% formalin with 0.1% Triton-X solution for intracellular staining?

A standard protocol for intracellular staining usually separate the fixation and permeabilization part. Can I do the fixation and permeabilization at one time by incubating cells with 3.7%...

17 July 2014 8,637 5 View

What is the difference between non-fat milk and BSA for primary antibody in western blot?

Some protocols mention primary antibodies should be diluted in 5%BSA whereas some protocols mention primary antibodies should be diluted in 5% non-fat milk. What is the difference? What is the...

26 May 2014 8,227 19 View

How to reduce cell clumping after cells have been treated with enzyme-free dissociation buffer?

Does anyone know how to reduce cell clumping after cells are treated with enzyme-free dissociation buffer? Is autoMACs rinsing solution (contains PBS and EDTA) helpful to prevent cell clumping in...

20 March 2014 8,096 8 View

Does anyone know the concentration of IL-2 that is sufficient to support resting NK cells to survive without activated to IL-2 NK cells?

see above

17 March 2014 6,087 0 View

Can anyone help me with these unusual results when multicolor staining in flow cytometry?

Currently, I am doing a research on NKG2D ligand expression on cancer cells. I would like to use dual color staining, which are APC-conjugated Ab for ULBP-1 and PE-conjugated Ab for ulbp-2....

04 March 2014 5,970 19 View