Anyone using addgene (www.addgene.org) to share plasmids?

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Trait introgression in plants: looking for ideas!

Dear all, I would like to hear ideas on how to speed up multiple trait introgression in plants. For example, introgression of multiple resistance traits or any other interesting trait. Anyone...

06 July 2017 408 0 View

How long can I store bacteria in LB to use it in a MiniPrep?

I was just wondering if it would be possible to store my bacteria after I grow them for a miniprep and what would be the best condition for it in case I just want to use them for a new DNA...

01 February 2016 3,856 5 View

Has someone already used TransTaq® DNA Polymerase High Fidelity for cDNA PCR amplification?

I would like to know if someone has used this Taq, TransTaq® DNA Polymerase High Fidelity, and if it compares with Phusion High fidelity. Thanks a lot!

31 December 2015 5,880 5 View

Help with stress gene evolution in plants!?

I need to prepare a talk on stress gene evolution in plants and I am having trouble finding material. This talk should be for undergrads... Anybody has something to share? Main topics I should...

08 September 2015 5,285 3 View

How to confirm the site-directed mutagenesis result without performing NGS?

I'm cloning a fragment of 3200 nts into plasmid. The cloning was successful, however, 02 amino acids were mutated. Now I want to fix these 02 aa by site-directed mutagenesis technique using...

08 August 2024 4,645 2 View

Where should the ARS sequence be introduced on a plasmid?

Hi all, I need to introduce an ARS (autonomously replicating sequence) in my plasmid but I'm not sure which position would be the best. Does anyone have any suggestion? A picture of the plasmid...

05 August 2024 1,573 4 View

Why after performing site directed mutagenesis ,I don't see any colony after transformation?

I want to introduce a point mutation (change in one nucleotide) into my gene of interest (DNA binding domain) I have designed primers as recommended on the Data sheet of the kit : -Both primers...

05 August 2024 9,059 3 View

Why my colony PCR results of my recombinant bacterial not showing any results?

I am performing ligation of the plasmid and a target gene. The steps I have taken are: 1. Double digestion of the plasmid and target gene 2. Ligation of the plasmid with the target gene 3....

05 August 2024 2,570 3 View

Transfection in HEK293T cells?

Dear All, I am trying to transfect a pCDNA3.1 vector containing my gene of interest. The purpose is to figure out the localization of the protein of interest. I have fused the protein with GFP on...

31 July 2024 9,892 4 View

Issues with Protein Expression in Transfected cells?

Hello I am trying to create a stable cell line in HEK293 via Lipofectamine 3000 transfection. My plasmid is a CD63-IL10-GFP construct with Puromycin resistance. I am successful with the...

30 July 2024 6,648 1 View

Does anyone have experience electroporating SKOV3 cells with a large plasmid using Neon transfection?

I have been trying to electroporate SKOV3 cells with a large plasmid (11kb) without much success. Any tips?

29 July 2024 3,229 1 View

Are these cassettes suitable for expressing PETase mutant in E. coli?

I created two potential gene expression cassettes (constitutive and inducible) for expression of a mutant PETase gene on PeptiCloud using the version tree feature, which allows users to create...

28 July 2024 7,559 1 View

What is the best way to drop a plasmid?

Please address the best way to drop a plasmid. Background: I have a "bait" plasmid resistant to kanamycin and a "prey" plasmid resistant to carbenicillin. After many rounds of streaking on...

25 July 2024 2,532 3 View

Isolation of PLV6-Bmal-Luc plasmid?

I am currently at a stage in my research where I will be using the pLV6-Bmal-luc plasmid (Addgene #: 68833). However, I am encountering some issues with the isolation of this plasmid and am unable...

23 July 2024 5,296 0 View