Flashcards
PCR
Cards in this deck (28)
- 1Anneal PrimersShort DNA sequences that define the area to be amplified
- 2Nestled primersBind inside the first set and are used for specificity
- 3Melt DNA at...95 degrees
- 4Primers bind after...Melting
- 5Anneal at...40-60 degrees
- 6Extend primers at...72 degrees
- 7The stages are repeated...20-30 times
- 8The ends of the amplified DNA representThe primers, not the actual DNA
- 9DNA Polymerases Vary InError Rate, Thermostability and DNA length required
- 10Uses of PCR- VirusesDiagnosis of Viral Infections
- 11Uses- CloningCreates millions of copies of a sequence very quickly
- 12Uses- DNAIndetification/ characterisation of a sequence
- 13Quirk of Taq PolymeraseA residues are added to the end of sequences
- 14T- VectorsAdd T residues to the ends to be ligated, which are complimentary to the A's left by Taq
- 15TOPO IsomeraseAn enzyme conjugated to the T/A residues, giving the vector a built in ligase
- 16Introduction of a Restricton SiteDone via the primer- use the site made to make insertion into the vector easier
- 17Reverse Transcriptase PCRConvert mRNA to DNA in PCR reaction
- 18Reverse Transcriptase PCR- PrimersCan be for PolyA tail or gene specific
- 19Reverse Transcriptase PCR- Tth Thermostable DNA PolymeraseHas both reverse transcriptae and helicase activity and can be used for both steps
- 20Quantitative PCRThe measuring of product as it is formed to determine how much DNA there was originally
- 21DNA Production in PCRSlows rapidly towards the end of the reaction
- 22Light Cycler Assay ProbesFluorescent beads that binds during amplification
- 23Light Cycler Assay- FluorescenceMore DNA produced, more fluorescence detected
- 24Use of Taqman AssayIdentify amounts of a specific DNA sequence
- 25Use of Taqman Assay ProbesContain fluorophore near a quencher
- 26Use of Taqman Assay AmplificationAmplification of DNA forces quencher of probe, causing fluorophore to fluoresce
- 27Molecular BeaconsProbe with a quencher and fluorophore- 2ndary strucuture of unbound probe holds them together
- 28Binding of Molecular ProbesPuts fluorophore & quencher far enough apart that fluorophore fluoresces
Information
- Description
- Year 2 Flashcards on PCR, created by gina_evans0312 on 29/12/2013.
- Created by
- gina_evans0312
- Views
- 560
- Tags
- Rating