Path: utzoo!utgpu!watserv1!watmath!uunet!bionet!hvrford.bitnet!J_MCLAUGHLIN From: J_MCLAUGHLIN@hvrford.bitnet Newsgroups: bionet.molbio.methds-reagnts Subject: Site-specific nucleotide base mutagenesis Message-ID: <9001051847.AA22125@genbank.bio.net> Date: 5 Jan 90 16:56:00 GMT Sender: daemon@genbank.BIO.NET Lines: 15 We are involved in making single base mutations in ribosomal RNA and then examinining how the mutations influence function. To make the mutations we have utilized or should I say attempted to utilize the Bio-Rad Muta-Gene kit. We continually have problems- I have checked all the reagents, my oligos and we still cannot get mutations. So, if anyone utilizing this method has some suggestions please let me know. Also, if you have any info on the E. coli strain- whether the dut ung efficiency inserting uracils is affected over time. Or, if anyone has some other "so-called" easy method to make mutations such as the T7 procedure from US Biochemical tell me PLEASE. The last thing I want to do is spend all of my time making mutations we want to see what they do! Thanks J_MCLAUGHLIN@HVRFORD