QuikChange II XL Site-Directed Mutagenesis Kit, 10 Rxn

Site Directed Mutagenesis (SDM) Human - Deletion PC-3 AGR2

Experiment
Site Directed Mutagenesis (SDM) Human - Deletion PC-3 AGR2
Product
QuikChange II XL Site-Directed Mutagenesis Kit, 10 Rxn from Agilent Technologies
Manufacturer
Agilent Technologies

Protocol tips

Protocol tips
- Use only the Dpn I enzyme provided; do not substitute with an enzyme from another source.

Publication protocol

Reporter vector construction and site‐directed mutagenesis
AR bound enhancer fragments of AGR2 and AGR3 were PCR amplified with the primers listed in Table 2 using Phusion high fidelity DNA polymerase (Finnzyme, Espoo, Finland) and inserted into the pSC‐B entry vector (Stratagene, La Jolla, CA, US). Inserts were size verified by KpnI and SacI restriction and sequencing, followed by ligation into a PGL3 promoter plasmid (Promega, Madison, WI, USA) using the Rapid DNA Ligation Kit (Roche) to generate firefly luciferase reporter vectors. Putative AREs were deleted using the QuikChange II Site‐Directed Mutagenesis Kit (Stratagene). The mutagenic primers used were 5′‐gctgAGCACAggaCACCGAgggaatggtgc‐3′ and 5′‐ggagtcAGGACAagcCACTCCtgctgaagtag‐3′ for AGR2 S1 ARE1 (AGCACAggaTGCTGA) and ARE2 (AGGACAagcTGATCC), respectively, and 5′‐ctgggtTGAGCTttgTTTGTCagagtcaactgtccc‐3′, 5′‐ccagagtCAACTGtccGTCGCCtgtactaaaatccacc‐3′ and 5′‐cagtctttaaAGTCTCagaTGACCAaggccttaaggtac‐3′ for AGR3 S1 ARE1–3 (TGAGCTttgTGTTCC, CAACTGtccCTGTCC and AGTACAagaTGACCA).

Full paper   Login or join for free to view the full paper.

Reviews

QuikChange II XL Site-Directed Mutagenesis Kit, 10 Rxn from Agilent Technologies has not yet been reviewed for this experiment

We'd love it if you would be the first to write a review!

Discussion

Start your discussion

Share your thoughts or question with experts in your field

Start a discussion

Papers

Check out relevant papers found by Labettor's AI that are relevant for performing Site Directed Mutagenesis (SDM) Human - Deletion PC-3 AGR2 using QuikChange II XL Site-Directed Mutagenesis Kit, 10 Rxn from Agilent Technologies.

Paper title
Anterior gradient 2 and 3--two prototype androgen-responsive genes transcriptionally upregulated by androgens and by oestrogens in prostate cancer cells.
Full paper
Login or join for free to view the full paper.

Manufacturer protocol

Download the product protocol from Agilent Technologies for QuikChange II XL Site-Directed Mutagenesis Kit, 10 Rxn below.

Download PDF Download manufacturer protocol

Videos

Check out videos that might be relevant for performing Site Directed Mutagenesis (SDM) Human - Deletion PC-3 AGR2 using QuikChange II XL Site-Directed Mutagenesis Kit, 10 Rxn from Agilent Technologies. Please note that these videos are representative and steps or experiment specific processes must be kept in mind to expect desired results.

We haven't found any additional videos for this experiment / product combination yet.

Outsource your experiment

Fill out your contact details and receive price quotes in your Inbox

  Outsource experiment
Become shareholder Discussions About us Contact Privacy Terms