PCR Flashcards
What are 3 examples of bioinformatics?
Manage data
retrieve and compare data
data mining & analysis
What is added to the end of mRNA to make it a mature mRNA?
Poly-A-tail
What is an ORF?
If RNA encodes a protein
comprises a stretch starting with ATG and ends with a stop codon
What are stop codons?
TGA
TAG
TAA
What is PCR?
exponential amplification of DNA sequences
What are 4 applications of PCR?
infectious diseases
forensics
genetic testing
molecular research
What are 4 PCR inputs?
Template - DNA containing target
Primers - oligonucleotides flank target
Polymerases - Taq
dNTPs - incorporation into product
What are primers?
Short - single stranded oligonucleotide
one pair per target (forward + reverse)
What are 6 primer factors to consider?
length
melting temp
annealing temp
GC content
GC clamp
3’ end stability
What are 5 primer pair factors?
Amplicon length
product position
Tm of product
optimum annealing temp
primer pair Tm mismatch calculation
What are 2 things to avoid in PCR primer design?
Primer secondary structures
cross homology, repeats and runs