Best way transfecting two sgRNA's on two different CRISPR/CAS9 plasmids?

I'm trying to knock out a gene in mESCs. I have my guides already cloned each into a CRISPR/CAS9 plasmid also carrying GFP. My guides didn't work after transfecting them individually, so I wish to...

03 November 2020 6,468 3 View

Which sensory-motor learning task and problem solving task are well-researched and easy to administer (do not require special & expensive equipment)?

We are planning a study in which we will explore the effects on the brain of a sensory-motor intervention which simulates the process of developmental sensory-motor learning. We plan to explore...

27 December 2018 6,935 1 View

Which resources could help residents develop a successful grassroots participatory process ?

In my article "When knowledge is power: grassroots participatory initiative as a process of resource development" I claim that residents are initiating informal grassroots participatory...

25 December 2017 1,657 12 View

Is slashing and mulching of Asparagus leaves at the end of the growing season more efficient than allowing nutrient translocation to take place?

In Australia, commercial asparagus farms cut back vegetative growth and mulch it into the soil at the end of the growing season in order to return the nutrients in these plant parts to the soil....

19 November 2017 988 3 View

Can any one give me a protocol for beta gal ONPG assay for AML12 transfected cells?

I have a protocol which we used a few years ago with 3T3 cells. With the AML12 cells I don't get any color. Any suggestions? Thank you.

30 October 2017 1,749 1 View

How to merge files and define new values on SPSS?

Well, I'm facing some troubles with SPSS. I'm trying to merge 2 files and I will try to explain the problem with a simple example: Let's say I have names of patients in the "master" file, while on...

14 September 2017 5,619 3 View

Which statistical model(s) is this paper using?

Differential tolerance to metals among populations of the introduced bryozoan Bugula neritina (Piola and Johnston 2006) I have been asked to review this paper and comment on the statistical models...

16 August 2017 9,131 3 View

Problem mixing the aqueous phase with isopropanol - RNAlater samples

Hallo everybody, I was extracting RNA (with TRIzol protocol) from bees in RNAlater. At the precipitation step we found that the aqueous phase was not mixing with the isopropanol - as if they were...

21 June 2017 6,379 1 View

Besides T7 RNA Polymerase, is there any other protein that binds the T7 promoter?

I know that T7 RNA polymerase does not require transcription factors , but is there any other protein besides the polymerase who can bind to the same promoter?

10 January 2017 8,049 2 View

What is the best solvent for extraction of 1,4-Butanediol diglycidyl ether (BDDE)?

18 October 2016 7,138 3 View

How can I measure MMP-1 protein activity or protein level from liver tissue?

Instead of checking only the mRNA level, I want to check the active protein level of MMP-1 in Liver tissue from mice. How can I do that?

03 March 2021 1,763 2 View

Does any one know how i can prepare liver tissue with transmission electron microscope with good fixation?

i have problem in preparation of liver tissue with bad cell organelles and how i can adjust osmolarity

03 March 2021 8,890 1 View

Interested to stain mice tissue with WGA 488 conjugate, anybody has a protocol to recommend?

Interested to stain mice brain tissue with WGA 488 conjugate, appreciate if anyone willing to offer me some guidance or suggestions! Thank you!

28 February 2021 3,951 2 View

Any specific pipeline or parameter to analyse total transcriptomics study of brain tissue samples?

I have two groups of brain samples, control and treated for example. It was total RNA nova seq sequencing. I tried all the available pipeline like: star+rsem+deseq2, Hista+stringtie+cuffdiff,...

27 February 2021 356 6 View

B-mercaptoethanol (BME) protocol?

Hi everybody, I have been running a multiplex IHC which allow me to stain the same tissue with different markers (Primary Ab + Secondary conjugated Ab) (AF487, AF555, AF647). Since I would like to...

27 February 2021 5,028 2 View

Why is the efficiency of viral mRNA electroporation in PBMC low?

Hello, I have been struggling with the electroporation of PBMC with HIV-I mRNA. Although the quality of the RNA generated is good, and the handling of the cells in meticulous, the frequency of...

27 February 2021 9,233 3 View

Looking for genomic DNA extraction kits from mouse tissue (centrifuge-based, not spin-column)?

Hello. I am looking to extract genomic DNA from mouse brain and lungs and I need a kit that is not spin-column based because I need to scale up the solution amounts for 400mg tissue (most cap at...

25 February 2021 155 2 View

Has anyone tried to scan fluorescence IHC slides with microarray scanner?

I have an old slide microarray scanner (VIDAR Revolution 4200) and would like to give it a "second life", as IHC scanner array (fluorescence dyes), FISH over tissue or similar. Has anyone tried...

24 February 2021 9,210 1 View

Recommendations on homogenizers for RNA isolation?

I will be performing RNA isolation with frozen lung biopsy samples and am in the market for a tissue homogenizer. I am wanting a homogenizer that will fit in a 1.5ml tube. I was looking at the...

24 February 2021 3,623 3 View

Is it efficient to do liquid biopsy to investigate the gene expressions of the tumor?

To do liquid biopsy and look for specific gene expressions there are so many published papers. Thera are different protocols and procedures used to isolate circulating cell free RNA or mRNA from...

23 February 2021 7,326 5 View