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

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

Has anybody worked on the detection of IGF1 protein in cell culture media?

Assay development

07 August 2013 6,115 2 View

How to repeat an operation on an atom selection for multiple times in PyMol?

Hi, I am trying to construct a multi-layer fibril structure from a single layer in PyMol by translating the layer along the fibril axis. For now, I am able to use the Translate command in PyMol...

02 March 2021 4,569 4 View

How to calculate average trend in maximum annual NDVI from MODIS?

I am wanting to calculate the average trend in maximum annual NDVI in Iceland from 2010-2020 using MODIS MYD13Q1 V6. How would I do this? I have currently inserted the NDVI bands from the MODIS...

02 March 2021 752 2 View

Why does two different bioinformatics tools for the same proteins show different subcellular localizations?

During a computational analysis, I found that for a protein of a plant, two different subcellular localizations are seen using CELLO and WoLF PSORT. For example, in CELLO, it identified the...

01 March 2021 6,729 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

What is the difference between genotype and haplotype data in the ped format?

Hi everybody In the ped format for genotype, alleles of any SNP are represented by two columns (one for each allele, separated by a space). Is a column sufficient for the haplotype to...

27 February 2021 1,965 1 View

I have need of XRD graph analysis software for phase identification. Can you please provide links for software tools which can be downloaded?

i am trying to download x'pert high score software, but can't download it. please do suggest software for phase analysis

24 February 2021 5,687 4 View

Please suggest me the simplest way to understand the in-silico analysis of any gene?

I want to understand the basics of docking, and also know about all bioinformatics tools which are free of coast and helpfull in in-silico analysis?

22 February 2021 8,879 2 View

Is there a Constitutive model for concrete with Sea-Sand?

Hi I am running a simulation in Abaqus for a concrete beam. Its fine aggregate replaced with sea-sand. I was wondering if I could find studies that measured the ultimate strain of concrete made...

22 February 2021 6,409 2 View

How can we measure the reliability and validity of our survey questionnaire?

We are conducting survey research on COVID-19 misinformation among students. How can we validate our survey for it to produce valid and reliable data? Also, what statistical tools can we use to...

16 February 2021 678 3 View

How to remove the species data from rda data frame in R (only remain the sites/samples point)?

Dear everyone, When I using the rda function in r, I met this question. As you can see in the figure, there are a lot of deep blue points which refers to the species. But I would like remain...

15 February 2021 8,982 2 View