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

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

What is the difference between RFP and dsRed?

When we construct plasmids, empty vectors containing RFP and dsRed were used usually, so what is the difference between each other?

04 May 2015 7,869 6 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

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

Gel electrophoresis result for total RNA samples, 1.5 Agarose gel was used, How to improve the integrity of RNA?

Gel electrophoresis, RNA degradation, RNA extraction from fresh tissue

02 March 2021 5,433 5 View

How to quantitative analize the data of TNBS assay for gelatin?

Estemeed colleagues, I found some issues regarding the quantification of the data for TNBS assay. There are different protocols on how to perform that but it is clear to me the "fil rouge" that...

02 March 2021 2,616 1 View

Does anyone know HRM channel in a 2 plex QIAGEN Rotor- Gene Q qPCR can be used as a separate channel?

I am going to have 3 different probes in my qPCR work that I am going to do. But I realized that the machine we have in the lab is a Rotor-Gene Q 2plex HRM Platform, saying it has green, yellow,...

01 March 2021 8,544 1 View

How to simultaneously perform multiple biochemical assays?

I am a research scholar working on heavy metal stress on plants. I will be doing biochemical characterisation( protein, carbohydrate, proline, antioxidant enzymes and many more assays) at interval...

01 March 2021 6,999 3 View

Which cell line should we use to perform biological dosimetry experiments?

We are preparing some experiments based on irradiating cells under different conditions in order to evaluate the effects in terms of DNA damage, genetic expression, etc. As our project is...

01 March 2021 3,355 3 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

Precipitate after adding laemmli buffer to protein solution containing Potassium Acetate?

Hi, I am running a size exclusion chromatography experiment with a buffer containing Potassium Acetate as a salt. I analyse these fractions through SDS-PAGE. After boiling my SEC fractions in...

01 March 2021 2,622 3 View

Is CD44 degraded in lysosome together with hyaluronan after CD44 mediated endocytosis of hyaluronic acid?

Also when RHAMM binds hyaluronic acid, they get internalized, will RHAMM also be degraded? Or both CD44 and RHAMM will be transferred back to the cell membrane? Asking for breast cancer cell line...

01 March 2021 8,169 2 View

What is the most suitable server for predicting protein structures in bacteria?

I have used the i-Tasser several times, however it has been unavailable for several days. I tried the swiss-model, but the output was not very pleasant due to the model used. Is there any other...

28 February 2021 4,521 3 View