How to clone the GOI into ROSA26 donor plasmid?

I am new to CRISPR Knock In. I want to clone a gene into ROSA 26 donor plasmid to Knock In a gene into the mouse genome. I have selected the addgene plasmid # 74285. But I cannot figure out where...

09 October 2019 9,506 1 View

Hello, I have a CRISPR related question. Can anyone help me?

I wanted to knock out MSH2 in mice cell line CH12F3 and after gRNA construction, transfection and single cell sorting, I got single colonies. I used two guide RNAs to give a deletion but when I...

04 May 2019 8,042 6 View

No title

for recombinant proteins do we have to use specific antibodies?

03 April 2019 9,757 3 View

How do I amplify a 7.4 KB fragment in mouse genomic DNA ?

I am struggling to amplify an 7.4 kB fragment which I need to confirm my CRISPR KO. I have tried Pfu and Phusion polymerase, used both HF and GC buffers, tried 50 - 70 C temp gradient, DMSO...

02 March 2019 1,361 4 View

How to design best primers to amplify a region in mice genomic DNA?

What is the database I should select under the Primer Pair specificity parameters in NCBI primer design tool?

02 March 2019 7,014 2 View

Anyone familiar with DBCO -CY5?

I need to purify DBCO CY5 since it give lots of unwanted bands when run on PAGE. I am looking for a protocol for that.

01 January 1970 239 5 View

Has anyone tried using zeta probe membrane in DNA slot blotting?

I do slot blotting to my DNA samples that contain Carboxy rhodamine tags and then scan through typhoon (CY2) to get the signal intensity. Unfortunately membrane looks terrible and I cannot...

01 January 1970 2,833 0 View

Can anyone suggest me a suitable solvent system for Cl and azide separation where both are linked to n-hydroxyphthalimide with an ethyl group?

Cl and azide linked to n-hydroxyphthalimide

01 January 1970 1,790 2 View

Can anyone help me with this Urea-PAGE gel issue?

When I run my 17mer DNA oligos on PAGE gel I can see a smear above my DNA band. Does anyone know a possible reason or how to minimize this? I have attached a gel image here as well. Thank you

01 January 1970 876 4 View

Can anyone help me with this question?

what is the most reasonable mechanism for hydrothiolation of terminal alkynes?

01 January 1970 7,687 0 View

MD MOE RMSD

How to calculate the RMSD values for a MD simulation using MOE?

07 August 2021 0 0 View

MD MOE RMSD

How to calculate the RMSD values for a MD simulation using MOE?

07 August 2021 0 0 View

fatal error

When I tried to energy minimization my system, I got fatal error as below. Fatal error: Atomtype opls_116 not found Although I've already added this line: ; include water #include "oplsaa.ff/spc.itp" to [molecultype] directive in my topology.

16 June 2021 0 0 View

fatal error

When I tried to energy minimization my system, I got fatal error as below. Fatal error: Atomtype opls_116 not found Although I've already added this line: ; include water #include "oplsaa.ff/spc.itp" to [molecultype] directive in my topology.

16 June 2021 0 0 View

fatal error

When I tried to energy minimization my system, I got fatal error as below. Fatal error: Atomtype opls_116 not found Although I've already added this line: ; include water #include "oplsaa.ff/spc.itp" to [molecultype] directive in my topology.

16 June 2021 0 0 View

fatal error

When I tried to energy minimization my system, I got fatal error as below. Fatal error: Atomtype opls_116 not found Although I've already added this line: ; include water #include "oplsaa.ff/spc.itp" to [molecultype] directive in my topology.

16 June 2021 0 0 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

What kind of fluid could I use for a pitfall trap that also does not invalidate molecular testing?

Hi, I want to start testing pitfall trap to obtain ants samples, but I need to conduct molecular analysis on those insects. So, what kind of fluid can I use? Ethanol expires too early and I need...

03 March 2021 5,978 5 View

No title

02 March 2021 3,060 3 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