Class Notes (808,312)
Canada (493,124)
Biochemistry (847)

Site Directed Mutagenesis

1 Page
Unlock Document

Western University
Biochemistry 2288A
Chris Brandl

Site Directed Mutagenesis Sometimes we want to slightly alter gene cause were not happy with wild typewe want to make direct changes Alter wild type to diff gene hopefully the gene will have different function How do we create a gene with altered sequence and in turn altered functionWe got our plasmid and we got normal gene inserted into it We have the codon that we want to change GAC codonFirst step create single stranded dna from it Separate two strands You can denature it or get virus to do itThen you synthetic oligonucleotidebut this oligo nucleotide has the specific base change in itit has GCC codon instead of GAC codonoligonucleotide carries that base change you want to incorporatein to the gene anneal this primer to single stranded dna and there is one mismatch at one position but it will steal anneal although not as good as wild type We then add dna polymerase and dntps and they will go around the plasmid making the strand duplex It will make dna duplex You also add dna ligase and it will fix the knick at the end We got
More Less

Related notes for Biochemistry 2288A

Log In


Don't have an account?

Join OneClass

Access over 10 million pages of study
documents for 1.3 million courses.

Sign up

Join to view


By registering, I agree to the Terms and Privacy Policies
Already have an account?
Just a few more details

So we can recommend you notes for your school.

Reset Password

Please enter below the email address you registered with and we will send you a link to reset your password.

Add your courses

Get notes from the top students in your class.