Molecular Genetics Flashcards
-Pentose, phosphate, base makes up
-RNA vs DNA sugar base
-DNA
-DNA has H, RNA has OH
Where does DNA replicate?
Where is RNA transcribed?
Where is RNA translated?
-nucleus
-nucleus; cytoplasm
-Protein K or magnetic beads used to
-Chloroform and Na acetate is what method
-extract DNA
-chemical
-Chaotropic salt
-ethanol
purifies DNA, protects NA
purifies DNA, precipates large biomolecules
-DNA is what charge?
-Ethidirum bromideis used as
-DNA is negatively charged, moves to anode
-dye in buffer
-larger the fragment, —-
-bands cut out from gel called
-to separate RNA, what do you use?
-slower it moves
-elution
-PAGE
dsDNA to ssDNA is called
nucleic acid hybridization
southern blot
-genome to ID sequence, electrophoresis, blot, blotted paper placed in bag to label it, rinse
PCR
-Taq DNA pol
-components
-synthesizes DNA or gene of interest using existing strand by adding dNTPs
-Taq, primers, dNTPs,buffer, DNA
-what specimen required for LAMP
-rtPRC
-saliva
-measures PCR product in each cycle
Traditional PCR vs rtPCR
-traditional = detection, DNA bands, primer increases
-rtPCR=probes or dyes (not primers), peak on graph (not bands), quantify it
-SYBR and TaqMan are
-rtPCR assays
SYBR = no sequence specificity
TaqMAn= high specificity
Which rtPCR is more specific?
TaqMAN
-removes RNA primers
-extends primers
-DNA pol I
-DNA pol II
autosomal codominant
blood group genes
-expressed when allele is present, antigen appears every generation
-expressed when one is homozygous for allele and inherits from parents
-girl chromosomes and boy
-autosomal dominance
-autosomal recessive
-XX
-XY
-father carries trait on X. All daughters will have the trait but not sons because sons get Y
-daughter will inherit the train in autosomal recessive way and sons will express it. when does this occur?
-X-linked
dominant
-if mom is X-linked trait, homozygouse for allele
-genes for different traits are inherited separately from each other
-two or more genes close to each other at loci tend to be inherited together. Do not assort independently. Two genes close to each are found together more times than chance alone
independent assortment
-linkage disequilibrium phenomenon
-3-5 direction is
-Which has less contamination: rtPCR or traditiona?
-template strand
-rtPCR
Reverse transcription performed by
is it RNA dependent or DNA
retroviruses
-RNA dependent DNA pol b/c makes DNA from RNA
-polymerase is inhibited by
-this requires more than one primer and detects more than two or more organisms
-heparin
-Multi-plex
-Which PCR is more sensitive
-amplifies signal
Nested
Branched
What do you put on Maldi-tof after colony dries out?
separated based on?
formic acid, laser will transfer protons to colony
mass to charge ratio
-western blot
-northern blot
-southern blot
-which part of gene is translated into a protein
-proteins
-RNA
-DNA
-exon
- How can false negative in PCR be detected
- most sensitive for viral meningitis: PCR or rtPCR?
- -what gene amplified in MRSA from CNS MRSA
-by using internal control
-rtPCR
-mecA/orfX gene
—— scatter is related to cell size and —– scatter to granularity
forward/side
-high frequency of mutation nucleotide
-cytosine
Facts
-Males can’t be heterozygous for X or Y since they only carry one copy. Hemizygous. Phenotypes are always expressed in them.
-maternal is mode of inheritance of mitochondrial DNA
-primers made using 16S subunit
-DNA more stable than RNA but RNA:RNA resistant to denaturation followed by RNA:DNA, DNA:DNA
—— joins nucleotides of DNA and hydrogen bonds between nitrogen bases
Phosphodiester bond
-adenine and guanines are
-uracil-N-glycolase used to contain
purines
contamination, removes uracil in DNA strands
homology
