Q5® Site-Directed Mutagenesis Kit

Site Directed Mutagenesis (SDM) Human - Deletion SKOV3 APRIN

Experiment
Site Directed Mutagenesis (SDM) Human - Deletion SKOV3 APRIN
Product
Q5® Site-Directed Mutagenesis Kit from New England BioLabs
Manufacturer
New England BioLabs

Protocol tips

Protocol tips
- Only use 1 μl of PCR product in the KLD reaction. Carrying too much PCR product forward can decrease transformation efficiency.

- Only use 5 μl of the KLD reaction in the transformation. If more KLD reaction is added, a buffer exchange step, such as PCR
purification, should be included prior to transformation.

Publication protocol

Constructs
FLAG/HIS tagged APRIN was cloned by polymerase chain reaction (PCR) amplification of pCMV SPORT plasmid APRIN-myc-his (gift from Peter Geck) into pFastBAC1-52b-GST using the primers cited in Table . pEGFP-APRIN was obtained after the insertion of EcoR1/BamH1 PCR products into pEGFP-C1 (Clontech). APRIN deletion constructs were obtained by mutagenesis using the Q5 Site-Directed Mutagenesis Kit (NEB) and primers cited in Table . APRIN fragments were cloned by PCR amplification of pCMV SPORT plasmid APRIN-myc-his (gift from Peter Geck) into pEGFP-C1 using primers cited in Table . mCherry-PCNA-19-NLS-4 was purchased from Addgene.

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Papers

Check out relevant papers found by Labettor's AI that are relevant for performing Site Directed Mutagenesis (SDM) Human - Deletion SKOV3 APRIN using Q5® Site-Directed Mutagenesis Kit from New England BioLabs.

Paper title
Roles for APRIN (PDS5B) in homologous recombination and in ovarian cancer prediction
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Manufacturer protocol

Download the product protocol from New England BioLabs for Q5® Site-Directed Mutagenesis Kit below.

Download PDF Download manufacturer protocol

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