Site-specific nucleotide base mutagenesis

J_MCLAUGHLIN at hvrford.bitnet J_MCLAUGHLIN at hvrford.bitnet
Fri Jan 5 11:56:00 EST 1990


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 at HVRFORD



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