How to improve the production efficiency of lentivirus?

Here, I am using HEK293T cells to product lentivirus. The plasmid system is pCDH, pMG-VSVG, pRSV-Rev and pMDLg-PRRL. I can get 1x 10^8 viruses from 10-cm dish HEK293T cells. It is not enough for...

08 September 2017 3,861 0 View

Is there any protocol of knocking out one gene in THP1 cells using CRISPR/Cas9 system ?

Is there any protocol of knocking out one gene in THP1 cells using CRISPR/Cas9 system ? Here, I used LentiCRISPR-v2 system to harvest virus carring guide RNA, but when I added the virus into THP1...

10 November 2016 6,267 2 View

Could HEK293T cells be used to package adenovirus?

We always use HEK293A cells to package adenvirus, could HEK293T cells be used to do it? Thank you~~~

08 September 2016 8,199 7 View

What is the function of SV40 poly(A) sequence in one plasmid? Does it regulate the gene of interest that is cloned into the plasmid?

Read 2 answers by scientists with 3 recommendations from their colleagues to the question asked by Yongxu Zhao on Sep 25, 2016

08 September 2016 9,128 2 View

How to prove one protein binding with DNA or RNA?

07 August 2016 3,146 0 View

In THP1 cells, how is a gene knocked out using CRISPR/Cas9?

As title, is there any great protocol to do this experiment?

01 February 2016 3,470 0 View

Which cells are similar with THP1 and are easier to be transfected?

01 February 2016 6,156 5 View

How is beta-catenin released from E-Cadherin?

09 October 2014 4,104 4 View

Why is the function of E-Cadherin totally different from N-Cadherin?

Both the structure and sequence of E-Cadherin are similar with those of N-Cadherin. However the function of these two cadherins is totally different. Why?

06 July 2014 142 9 View

How to coculture macrophage and cancer cell?

Does anyone have a detailed protocol for macrophage and cancer cell cocultures?

10 November 2013 9,184 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

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

Did anyone ever try using pLVX-TetOne Plasmid with a copGFP as a selection marker instead of Puromycin?

When using a lentiviral vector for inducible expression of genes using the Tet-On system. In the literature, I saw a widely used of pLVX-TetOne-Puro Plasmid, is there a reason why people prefer to...

25 February 2021 1,011 3 View

Why do the PCR bands from the cDNA templates (extraction from the cell after transfection) show different size from the band of the plasmid template?

After transfection with the plasmid ( linearized ) and subcloning of the cell lines, RNA was extracted from the cell and then reverse transcripted to cDNA. When PCR reactions were run to verify...

25 February 2021 5,712 3 View

Does anyone have a sequence for the human CD34 promoter?

I'm looking for the human CD34 promoter sequence to engineer into a vector construct. Can anyone recommend a good source for this sequence? Thanks very much, Laura

25 February 2021 8,205 1 View

How do I transfect a large plasmid into PC12 cells with nerve growth factor (NGF) receptor?

I have a large 16 kb plasmid, which I need to transfect into PC12 cells. Lipofectamin 2000 didn't work and with GFP alone the transfection rate is very low. I also tried the neon invitrogen...

25 February 2021 6,635 5 View