Macrophage/Microglia from PBMC?

Hi, I am trying to isolate monocyte from PBMC and induce them to make macrophages and microglia. I used monocyte attachment media and then change after 2 hours to media required for macrophage...

08 September 2019 1,187 4 View

LncRNA absent from mouse genome?

Hi, I am working with a lncRNA (uncharacterized ) which i found to be present within the intonic region of my gene in human. When i blast it with mouse genome i did not find any sequence...

03 April 2019 3,628 3 View

Can i express lncRNA and protein using IRES sequence?

Hi I am trying to express my lncRNA in neural cells. As transfection efficiency will be low i am planing to use lentivirus as my delivery method. But to be sure can i add a GFP protein using IRES...

02 March 2019 1,744 9 View

Mouse B cell immunology??

Hi All, I am trying to access changes in B cell markers in our knockout mouse and wild type from blood. I try to gate lymphocytes, then single cell, followed by live dead and B220+ cells. When i...

01 February 2019 3,960 2 View

Transfection problem in HELA CELL???

Hi , i tried many transfection reagent and Viafect worked best for me when i used in HELA cell (knockdown for some protein) as observed by reporter plasmid (5.4 kb). When i do my experiments i...

04 May 2018 8,693 7 View

Can anyone help me analyse my RT-PCR data?

01 February 2016 7,751 4 View

RIN no for exosome microRNA analysis?

11 December 2015 9,198 4 View

Enhanced Yellow Fluorescent Protein (Aequorea victoria) DNA sequence?

I am on the lookout for the Enhanced Yellow Fluorescent Protein (Aequorea victoria) DNA sequence. Does anyone know where I can find it? Thank you in advance

03 March 2021 3,568 1 View

Which is a lucid standard comprehensive book to guide on psychological scale construction and its standardization?

02 March 2021 7,082 3 View

Does anyone have a recommendation for yeast reporter gene plasmid/protocol?

Hi, could anyone recommend a plasmid and/or protocol for reporter gene assay in S. cerevisiae? I want to assess the effect of growth conditions on a transcription factor, so I want to clone it`s...

01 March 2021 210 1 View

Why might my GFP tag be dissociating from my plasmid after transfection?

I have a set of stably transfected cell lines all transfected with plasmids containing GFP tags on the C terminus. During a western blot using anti-GFP antibody, one of my plasmids has dissociated...

01 March 2021 9,310 4 View

Is it possible for two human neuronal cell lines to have different gene sequence for the same protein?

I am looking at the ATP1A2 (Sodium/Potassium ATPase alpha subunit 2) in two human neuronal cell lines. Expression levels of this protein seems to be almost equal when detected by one antibody....

01 March 2021 3,607 3 View

Can someone suggest a way to prepare slides to observe under confocal microscope for observing fluorescent tagged proteins??

I am growing the cells on coverslips and transfecting the fluorescent tagged protein directly on them. Then i want to observe these cells under confocal microscope. Currently i am just using...

01 March 2021 6,142 3 View

Expression cloning by using cDNA,do I need to modify the cDNA first?

I am going to have a expression cloning of mammalian gene by using shuttle plasmid to transforming the E.coli However I don't know I should only inserting the Coding sequence ,or I can...

28 February 2021 5,440 3 View

Problem with shRNA transfection. Why are only two plates getting contaminated instead of all?

I transfected my LNCaP-WT cells with 3 shRNA plus their NTC two weeks ago and split two puromycin selected cell plates on Friday last week(Feb 26). I checked for GFP in the cells, and they all...

28 February 2021 4,949 3 View

PUC19 as a transfer plasmid for lentiviral delivery?

Hello, Is it possible to use pUC19 as a transfer vector to be packed in using the second generation viral particles packaging system( pMD2.G; psPAX2 plasmids)? As far as I understand it there is...

28 February 2021 4,868 2 View

Runs of nucleotides in Sanger sequencing?

I performed site directed mutagenesis, transformation, and then I sent out plasmids for Sanger sequencing and found out that there is extension of DNA just before the stop codon. I am not sure...

27 February 2021 547 3 View