Lecture 1 and 2 - Technologies Used in Genetics
Heather Knight
Polymerase Chain Reaction - Method of Amplifying DNA
Measuring Gene Expression - Methods for Quantifying mRNA Levels (num of transcripts)
Technologies used in Genetics with regard to DNA and RNA analysis
The many applications of PCR is to measure Gene Expression
PCR
• Method of amplifying DNA
Amplified DNA —> Cloning and Detection/Characterisation
Why want to Amplify DNA ?
• To make enough DNA to clone or want to produce enough DNA to detect presence of particular piece of
DNA and/or to characterise it efficiently
• Usually not enough DNA thus need to amplify more
• Makes lots of copies of DNA, dominant genes would be abundantly present
• If want to analyse 1 gene amongst a genome containing thousands of diff genes, it’s diluted out by all other
uninterested thousands of genes
• If amplify 1 gene interested in, sequence is enriched and easier to examine or use in other applications
PCR - 3 Steps
1. Denaturation
2. Annealing
3. Extension
* Process repeated numerous times
• Red part interested DNA
• Exponential doubling of DNA
• 1 copy of DNA converted to many copies
• Doubling of amount of material in each cycle
1. Denaturation
Double stranded DNA —> 94~98 degrees —> 2 Separated strands (single strand DNA)
At very high temp, double stranded DNA separated into 2 separate strands
PCR method usually use temp in region of 94-98
Heather Knight
Polymerase Chain Reaction - Method of Amplifying DNA
Measuring Gene Expression - Methods for Quantifying mRNA Levels (num of transcripts)
Technologies used in Genetics with regard to DNA and RNA analysis
The many applications of PCR is to measure Gene Expression
PCR
• Method of amplifying DNA
Amplified DNA —> Cloning and Detection/Characterisation
Why want to Amplify DNA ?
• To make enough DNA to clone or want to produce enough DNA to detect presence of particular piece of
DNA and/or to characterise it efficiently
• Usually not enough DNA thus need to amplify more
• Makes lots of copies of DNA, dominant genes would be abundantly present
• If want to analyse 1 gene amongst a genome containing thousands of diff genes, it’s diluted out by all other
uninterested thousands of genes
• If amplify 1 gene interested in, sequence is enriched and easier to examine or use in other applications
PCR - 3 Steps
1. Denaturation
2. Annealing
3. Extension
* Process repeated numerous times
• Red part interested DNA
• Exponential doubling of DNA
• 1 copy of DNA converted to many copies
• Doubling of amount of material in each cycle
1. Denaturation
Double stranded DNA —> 94~98 degrees —> 2 Separated strands (single strand DNA)
At very high temp, double stranded DNA separated into 2 separate strands
PCR method usually use temp in region of 94-98
