What amount of construct DNA should I use in the nucleofection protocol for CRISPR-Cas9 gRNA?

I want to perform the nucleofection of CRISPR/Cas9(WT) gRNA construct in beta cell line. 1. How much construct DNA shall I use for the nucleofection. Technique- Amaxa Nucleofector Kit- T or Kit-...

04 May 2015 2,133 5 View

How do I solve my problem in generating Bacmid?

04 May 2015 8,670 4 View

What Bioinformatics tool can I use for PCR efficiency check ?

I am looking for tool to evaluate the PCR efficiency for any template/gene. Especially to get an idea about the nature/complexity of gene stretch e.g. Any stable secondary structures, polymerase...

03 April 2015 2,313 4 View

How can I digest only one RE site of vector if having two sites of the same REs?

I am using pIRES bicistronic vector for cloning, where it contains two sites of Sma-I in MCS-B region. I have designed cloning primers with sma-I forward and Sal-I reverse. Since another sma-I...

02 March 2015 525 10 View

Concept of Bed volume in protein purification ?

Please explain the concept of bed volume in protein purification and how to calculate it ? E.g. I am using glass Econo Column(1.0X10cm)Biorad#737-4011 and using Ni-NTA slurry of 0.5 ml or 1mL....

01 February 2015 6,483 1 View

How can I improve the purity and quantity of his-tagged protein?

I am purifying one of the tyrosine kinase protein expressed as his-tagged in SF-9 cells, however I am facing problems in Ni-NTA protein purification. The details of the procedure is as...

11 December 2014 1,472 16 View

What is the ideal lysis buffer composition for colon cells like FHC and CCD-18Co?

I tried buffer: 1%NP40, 20mM Tris, 137mM Nacl, 10%glycerol, 2mM EDTA, 1mM activated Na-Orthovanadate, Aprotinin 10ug/ml,leupeptin 10ug/ml. Incubation with lysis buffer for 30min but still I see...

03 April 2014 8,302 4 View

How can I perform a search in order to find out the promoter region of any gene?

eg. if I want to search the promoter for IGF1 gene.

02 March 2014 4,212 4 View

Can anyone guide me on how to search for the promoter region of any gene??

Molecular biology

02 March 2014 5,338 5 View

After a reference from Int J Body Compos Res, can any body help?

Hi, I am after the reference below, my library says it cannot obtain a copy either locally or internationally, any help appreciated! Chris Wang ZM, Heshka S, Wielopolski L, Pi-Sunyer FX, Pierson...

03 March 2021 6,193 1 View

No title

02 March 2021 3,060 3 View

New Journal, "Integrative Psychological and Behavioral Science" -- do you not know, and have you not seen, this done before?

Question to you and THEM, the New Journal, "Integrative Psychological and Behavioral Science" -- do you not know, and have you not seen, this done before? There appears to be a core problem for...

02 March 2021 3,024 2 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

No title

01 March 2021 3,905 5 View

Issues with Permeabilization during Flow Cytometry?

Hello Everyone. Currently I am working to characterize macrophages in the myocardium after ischemia-reperfusion injury in rats. Due to the low total cell number isolated from rat hearts I can...

01 March 2021 3,867 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

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

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

Do you need the CMV enhancer before the CMV promotor for a proper function of CMV on the gene that follows?

I'm a student and I have to produce a cell line with a knockin for the NRF2 gene with GFP. I have to put a promotor in front of the GFP because the gene will be too far away from the promotor of...

28 February 2021 7,127 2 View