The expression plasmids, including pCMV-F-NTPase, pCMV-F-P22, and pCMV-F-Nterm, have been described previously (15 (link)). The plasmids pCMV-NTPase-GFP and pCMV-NTPase(1–179)-GFP were constructed by inserting the coding regions of NTPase and NTPase(1–179), respectively, into pEGFP-N2 (6081-1, Clontech). To obtain the series of the N- or C-terminal deletions of NTPase(1–179) in pCMV-NTPase(1–179)-GFP, the corresponding DNA fragments were amplified by PCR and then cloned into pEGFP-N2. The mutant constructs that contain internal deletions in pCMV-F-NTPase and pCMV-NTPase(1–179)-GFP were generated by using inverse PCR. Point mutations in pCMV-NTPase(1–50)-GFP or pCMV-NTPase(51–90)-GFP were created using a QuickChang site-directed mutagenesis kit (200524, Agilent Technologies). The plasmid that encodes NTPase(1–90)-RFP was constructed by inserting the RFP-coding fragment downstream of the NTPase(1–90)-coding sequences in pFLAG-CMV-2 (E7398, Sigma-Aldrich).
Quickchang site directed mutagenesis kit
Manufactured by Agilent Technologies
The QuickChange site-directed mutagenesis kit is a laboratory tool designed for introducing specific mutations into double-stranded plasmid DNA. The kit provides a rapid and efficient method for creating point mutations, insertions, or deletions within a DNA sequence.
Automatically generated - may contain errors
Lab products found in correlation
2 protocols using quickchang site directed mutagenesis kit
1
Cloning and Characterization of NTPase Constructs
2
Cloning and Characterization of NTPase Constructs
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The expression plasmids, including pCMV-F-NTPase, pCMV-F-P22, and pCMV-F-Nterm, have been described previously (15 (link)). The plasmids pCMV-NTPase-GFP and pCMV-NTPase(1–179)-GFP were constructed by inserting the coding regions of NTPase and NTPase(1–179), respectively, into pEGFP-N2 (6081-1, Clontech). To obtain the series of the N- or C-terminal deletions of NTPase(1–179) in pCMV-NTPase(1–179)-GFP, the corresponding DNA fragments were amplified by PCR and then cloned into pEGFP-N2. The mutant constructs that contain internal deletions in pCMV-F-NTPase and pCMV-NTPase(1–179)-GFP were generated by using inverse PCR. Point mutations in pCMV-NTPase(1–50)-GFP or pCMV-NTPase(51–90)-GFP were created using a QuickChang site-directed mutagenesis kit (200524, Agilent Technologies). The plasmid that encodes NTPase(1–90)-RFP was constructed by inserting the RFP-coding fragment downstream of the NTPase(1–90)-coding sequences in pFLAG-CMV-2 (E7398, Sigma-Aldrich).
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!