Lab Techniques Flashcards
Genome
Total genetic content of an organism
Transcriptome
Full complement of unique sequenced RNA transcripts that an individual produces; includes mRNA, rRNA, and tRNA
Proteome
Entire set of proteins expressed by a genome
Western Blot
Detects protein
Specific proteins can be detected with antibody labeling
Southern Blot
Detects DNA
Northern Blot
Detects RNA
SDS-Polyacrylamide Gel Electrophoresis (PAGE)
Since proteins don’t have a uniform charge, can heat them with SDS and Mercopteoethanol to get uniform charge then run electrophoresis
Polyclonal Antibody
Mixture of different antibodies targeted to different epitope sites
Epitope site
The part of an antigen molecule to which an antibody attaches itself
Monoclonal Antibody
Prepared to react against a single epitope site on a protein
Type II Restriction Endonucleases
Cleave DNA within their recognition sequence. Can either make “stick ends” with overhanging ends of DNA or “blunt ends” with flat ends of DNA at the cut site
Palindrome
When read from the 5’ to 3’ direction, the sequence on the “top” strand is identical to that of the “bottom” strand
Mass Spectrometry
Used for analyzing proteins and mass-to-charge ratio of ions. Gives protein abundance and peptide sequence.
Elisa
Used for detecting and analyzing proteins and antibodies
qPCR
Used for analyzing DNA and RNA
Sanger sequencing
Can give us the sequence of DNA and RNA
Next Generation Sequencing
Can give us the sequence of DNA and RNA
How are enzyme Km and enzyme/substrate affinity related?
They are inversely related. A higher Km means a lower enzyme/substrate affinity, and vice versa.
Microarray
Determines the mRNA production (gene expression) from thousands of genes at once
