RNA Isolation
What is the function of DEPC (diethyl pyrocarbonate) in RNA isolation?
It inactivates RNases.
Flow cytometry is routinely used diagnostically to evaluate what kind of diseases?
Leukemias and Lymphomas
Nucleosides are made up of a ribose sugar and a nitrogen base. True or False?
True
The most commonly used electrophoresis method in Western Blotting is called?
SDS-PAGE
What is the general term used to refer to all post-Sanger sequencing technologies?
Next-generation sequencing (NGS)
What reagent is used in the organic isolation of total RNA?
Trizol
In flow cytometry, what do the forward scatter and side scatter measure?
Forward scatter: cell size
Side scatter: internal complexity (granularity)
What are the two main pyrimidine ring structures found in DNA?
Thymine and Cytosine
List three samples used for Western Blotting.
Whole tissue, cell lysates, serum or body fluids, bacteria, viruses, and environmental samples.
Pyrosequencing involves the synthesis of DNA by the addition of nucleotides. True or False?
True
In RNA isolation using Trizol, what phase contains the RNA?
Aqueous phase
List the three components of a flow cytometer
Fluidics system, Optical system, Electronics
Which enzyme cuts DNA strands internally?
Endonucleases
List the steps involved in Western Blotting from steps 1 to 6.
Sample preparation, Gel electrophoresis, Transfer of bands to a membrane, Blocking, Detection, and Analysis.
What inhibits strand extension during Sanger Sequencing?
Dideoxynucleotide triphosphates (ddNTPs)
List 3 sources of RNA
Whole blood, Tissues, Cultured cells.
List the four major types of ELISA
Direct ELISA, Indirect ELISA, Sandwich ELISA, Competitive ELISA
List the two Non-coding RNAs.
Ribosomal RNA (rRNA)
Transfer RNA (tRNA)
What happens in step 1 of Western Blotting?
The Sample is lysed to release the proteins and treated with SDS.
In Sanger sequencing, what enzyme synthesizes DNA fragments?
DNA polymerase
Calculate the RNA concentration in μg/mL from the following information:
Absorbance reading at 260nm from a 1:100 dilution = 0.162
0.162 x 40μg/mL x 100 = 648 μg/mL
What ELISA method did we perform in this laboratory, and what did we detect in the patient sample?
Indirect ELISA, Antibodies
Which protein structure involves interactions between multiple polypeptide chains?
Quaternary Structure
Why is blocking done in step 4 of Western Blotting?
What are the two commonly used blocking agents in this step?
To prevent any non-specific interactions of antibodies and proteins.
Bovine serum albumin (BSA) or Non-fat dry milk.
List three processes involved in the Next-generation sequencing technique.
Library preparation, clonal amplification, cyclic array sequencing.