Debosree Pal

9 Questions 28 Answers 0 Followers

Questions related from Debosree Pal

Can anyone help me with culture and transfection of primary neurospheres?

Hi, I am growing neurospheres from progenitors isolated from the cortical region of E14.5 mouse brain. I want to perform sh/siRNA mediated knockdown of my gene of interest in the neurospheres....

05 December 2017 4,528 5 View

How to understand if homozygous knockout is lethal during Crispr mediated knockout generation?

Hi, I am trying to generate a homozygous knockout line for a particular gene in mouse embryonic stem cells by CRISPR/Cas9 mediated recombination. My strategy is to replace my target gene with GFP...

10 June 2017 7,248 20 View

Has anyone worked on embryonic germ cells as a model system?

I want to work on embryonic germ cells. Can anyone suggest from where I can procure these cells? Also, does anyone know the difference between embryonic germ cells and primordial germ cells

24 February 2015 5,247 1 View

Why do cells keep dying after two passages even if ther's no mycoplasma contamination?

I am working on mouse embryonic stem cells (R1) and have been facing a lot of problems of late in its culture. After reviving, the cells grow fine on feeders for about 2passages. In the third...

20 December 2014 2,055 6 View

Can R1 embryonic stem cells be grown in 2i+LIF medium?

Hello, I am growing R1-ESCs in high glucose DMEM, 15%FBS, sodium pyruvate, non-essential amino acids and L-glutamine. Can these cells be grown in 2i+LIF medium? How many passages do I need to grow...

23 October 2014 6,583 4 View

Why are mouse embryonic stem cells not growing properly?

I am working with the R1-ESC line. I was growing them on feeders on mitomycin C inactivated MEFs using FBS from Invitrogen (South American Origin). They were growing fine in this FBS. Recently,...

20 August 2014 4,395 6 View

What are the criteria for designing probes for whole embryo in situ hybridization?

I am doing in situ hybrdization on mouse embryos for a novel gene and so have to include both antisense and sense probes. The problem is I get the same staining pattern for both the probes. I have...

03 July 2014 7,894 17 View

Can anyone help me with mitomycin C inactivation of fibroblasts?

I have isolated primary mouse embryonic fibroblasts from CD-1 mice. They grow fine until P4/P5. Generally at P3, I arrest them for use as feeders. I use mitomycin C at a final concentration in the...

17 May 2014 2,675 13 View

Does the method of homogenization of tissues bring about a change in downstream PCR results?

I was trying to see the expression of a gene in post-implantation mouse embryos from 9.5-13.5dpc. The preferred technique is obviously whole embryo in-situ hybridization but, I first thought of...

01 May 2014 6,040 3 View