RNA Extraction: Sample and purify mRNA from

pigs
Question 1: What word fits in the gap?

When a gene is expressed ... is produced.

✓ mRNA
– siRNA
– a signal
– a complementary DNA strand

Question 2: All genes are expressed in all the cells. Is this statement true?
✓ No, only genes that are relevant for that particular cell are expressed
– No, only half of them are expressed in all the cells
– No, all genes are only expressed in the young cells and then the expression level decreases
– That's true

Question 3: If mRNAM R N Ay from gene A is very abundant in the fat tissue from the obese pig
and rare in the fat tissue from the lean pig it shows us that .... ?
✓ Gene A is more expressed in obese animals
– No conclusion can be drawn from this data
– Gene expression is random
– Gene A is less expressed in obese animals

Question 4: Why do we keep tissue samples for RNA work in liquid nitrogen (-196oC)?
✓ To preserve the RNA, which easily degrades at room temperature
– To deactivate the RNA
– To preserve the tissue structure
– To keep the fat tissue from melting

Question 5: The RNA content in a cell ....

✓ Includes both coding mRNA and several types of non-coding RNA
– Includes only coding mRNA
– Includes only mRNA and tRNA
– Is restricted to mRNA

Question 6: Before we start, do you know where is most of the mRNAM R N Ay located?
✓ In the cytoplasm
– In the nucleus
– In the cellular membrane
– In the mitochondria

Question 7: The lysis buffer used for RNA extraction contains ...
✓ Guanidinium isothiocyanate and phenol
– None of these answers
– Isopropanol
– Chloroform

Question 8: What does the lysate consist of?

✓ RNA, DNA, proteins and cellular debris
– Intact cells
– RNA and proteins

Page 1
 – RNA, DNA, proteins

Question 9: What do you measure at 230, 260 and 280 nm, respectively?
✓ Chemicals, RNA/DNA and proteins
– RNA at all wavelength
– Proteins, DNA and RNA
– DNA, proteins and RNA

Question 10: A 260/280260 to 280 ratio should be between 1.8 and 2.1. What does it mean if we
have a 260/280260 to 280 ratio that is lower than 1.8?
✓ There are a lot of proteins in the RNA sample
– The RNA sample is very clean
– The RNA sample is very degraded
– There are some chemical remnants e.g. phenol in the RNA sample

Question 11: What will a spectrophotometer measure?

✓ Purity and quantity
– Purity and integrity
– Only purity
– Purity, integrity and quantity

Question 12: What should you see in the gel if the RNA is intact?
✓ Two bands corresponding to ribosomal RNAs (rRNAs)
– A smear
– One band corresponding to all mRNAs
– Two bands corresponding to ribosomal RNas (rRNAs) and a third band corresponding to
mRNAs

Question 13: Why do we want to specifically extract the mRNAM R N Ay only from the total RNA?
✓ Because mRNA represents the genes that are expressed and will be translated into proteins
– Because mRNA represents all the other types of RNA
– Because mRNA is located only in the nucleus
– Because mRNA is the most abundant RNA and hence it is easy to sequence

Question 14: What is the common sequence that all mRNAM R N Ay share?
✓ polyA
– polyG
– polyT
– polyC

Page 2