Quiz 1 PPT Notes Flashcards
Why clone and make GFP-fusion proteins when antibodies localize the protein the cell?
May not …
May want to look at …, … cells
have antibodies;
dynamic; living
Using antibodies gives us great data on … But, we need to make antibodies (takes …, ….)
localization;
time;
variable recognition
what gives cells their shape? the cytoskeleton
- cellular “skeleton” that gives the cell structure and shape
- serves as tracks for … proteins
- participates in … and …
- drives …
- also has roles in …
- cytoskeletal proteins are …
motor; mitosis; cytokinesis; cell migration; cell adhesion; highly conserved
Three cytoskeletal systems/elements:
…
…
…
microtubules;
intermediate filaments;
actin filaments
stages of first 75 hr of sea urchin embryo. notice many changes:
1. number of … –> …
2. change in … shape, change in … shape
3. … of cells
(all things that rely on the …)
cells; cell division;
embryo; cell;
movement;
cytoskeleton
DNA nucleotide =
… + … (…) + …
phosphate group;
sugar; deoxyribose;
base
DNA:
phosphate backbone = … joins …
5’ phosphate;
3’ OH
DNA is: double stranded held together by H bonds ... strands double helix all of these contribute to increased stability, which is exactly what we want for something that is responsible for genetic info
antiparallel
how do we manipulate DNA?
- … it - …
- … it - …
- … it
amplify; PCR;
digest; restriction enzymes;
sequence it
how do we manipulate DNA?
- … it - … into …
- … it - ….
- … it - ….
clone; ligate; vectors;
label; hybridization;
analyze; agarose gel electrophoresis
nucleotide entry tells us different sequence features:
- …
- ….
- … features
- … sequence
- … sequence
source; gene length; coding sequence; protein; mRNA
Determine the … by finding the start and stop codons by eye or by using a translation tool
open reading frame (ORF)
the absorbance for a molar concentration of a substance with a path length of / cm determined at a specific … Its value is obtained from the equation …
wavelength;
epsilon = A/cl
beer lambert equation: …
A = epsilon * b *c
beer lambert equation: A = epsilon * b * c A: ... epsilon: ... b: ... c: ...
absorbance;
molar absorption coefficient;
path length;
concentration
In DNA, … can be deaminated and become uracil. There are enzymes that recognize the G-U mismatched base pairs, excise out the uracil, and replace with a …, thereby maintaining the accuracy of the genetic material
cytosine;
cytosine
rRNA: … RNA, is made in the … of the …, and transported to the cytoplasm where they will form ribosomes
ribosomal;
nucleoli;
nucleus
tRNA: … RNA, base pairs with the codon sequence on mRNA and brings in the appropriate amino acid during translation
snRNA: …, reside in the …, participate in …
transfer;
small nuclear RNAs; nucleus; splicing
snoRNA: …, reside in … and participate in …
small nucleolar RNAs;
nucleoli;
editing rRNA
… and … RNAs act at the level of … regulation, controlling the translation of the mRNA into protein
microRNA;
silencing;
posttranscriptional
other species of small RNAs are involved in regulation of … and …
transcription;
translation
how do we isolate RNA:
… cells –> inhibit … activity –> … –> recover intact RNA
lyse;
ribonuclease;
remove proteins
what can be done with isolated RNA:
- …. (aka …)
- …
- …
- … and …
- …
- ….
northern analysis; northern blots; nuclease protection assay; in vitro translation; cDNA library construction; screening; RT-PCR; qRT-PCR
Northern analysis is a method used to determine:
… of an mRNA
… of transcript
presence of …
abundance;
size;
alternative splicing
cDNA library:
- a collection of all the … at that time in that tissue/cell
- isolated mRNA is reverse transcribed into cDNA
- the cDNA is digested with restriction enzymes and …
- the library can be amplified in … and screened for specific sequences
expressed gene sequences;
ligated into vectors;
bacteria;
Going from RNA to DNA occurs in:
…
…
…
retroviruses;
retroposons;
telomeres
Reverse transcriptase is a …
RNA-dependent DNA polymerase
for reverse transcription we need: ... ... ... ....
RNA template;
primer;
reverse transcriptase;
nucleotides
+RT is a …
positive control
for - RT: since no RT in reaction, no … can be made. Therefore, any product in PCR is due to … contamination
For -RNA, since there’s no RNA in the reaction, nothing is there to provide a … Therefore, any product in PCR is due to one of the reaction components being contaminated
cDNA;
gDNA;
template
forward primer has … and is the …
reverse primer for actin has … To form reverse primer … and …
ATG; direct sequence;
TTA; reverse; complement;
Location of primer depends on …, aka what you want to …
what you want to do;
copy
