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molecular genetics > exam/paper questions > Flashcards

exam/paper questions Flashcards

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1
Q

Do you need to round the amount of template DNA in determining volumes for PCR?

A

No!

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2
Q

During denaturation what is the temperature used and the state of DNA?

A

Temperature- 94 or 95ºC
DNA- single stranded

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3
Q

During annealing what is the temperature used and the state of DNA?

A

Temperature- 52-65ºC
DNA- ds/ss hybrid

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4
Q

During elongation what is the temperature used and the state of DNA?

A

Temperature- 72ºC
DNA- double stranded

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5
Q

When performing dilutions what glassware do you use for mixing/dissolving?

A

Beaker or e. flask

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6
Q

When performing dilutions which glassware do you use to fill to final volume?

A

Graduates cylinder

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7
Q

Do you need to round GC content?

A

No

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8
Q

What is the function of the positive control in PCR?

A

To ensure that the assay components are working, tap, primers.

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9
Q

If you had a restriction enzyme that cut at 2 positions how many bands would present on homozygous D/R and heterozygous individuals in PCR?

A

Homozygous dominant- both strands have cut positions - 3 bands
Homozygous recessive- neither strand has a cut position - 1 band
Heterozygous- one band has cut positions, other doesn’t - 4 bands

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10
Q

What are two consequences of increasing degeneracy in primers?

A

Increased chance of non specific binding and a decrease in overall amplification

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11
Q

What are three consequences of having a run of the same nucleotide longer than 4 bases?

A

Increase secondary structures, increase non specific binding, increase mis-priming

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12
Q

What are two potential reasons as to why a PCR would not work?

A

Human error and remaining protein or secondary metabolite contamination

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13
Q

Why did Wendell et al., 2016 authors need to “further refine the position of the anl gene”? What other evidence did they use to confirm this?

A

It is an inverted repeat making it difficult to use insert specific primers on since amplification is not possible. Used phenotypic evidence from other plants.

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14
Q

How do you determine the most probably ancestral amino acid sequence using parsimony?

A

Looking at all sequences and seeing which amino acid is most common

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15
Q

What is the equation used for chi squared analysis?

A

(O-E)^2/E

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16
Q

How did rbcL contribute to universality and discrimination?

A

okay discrimination and high universality

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17
Q

How did matK contribute to universality and discrimination?

A

high discrimination, mixed universality

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18
Q

How did trn-psbA intergenic spacer region contribute to universality and discrimination?

A

high discrimination, high sequence variability

19
Q

How did atpF-H contribute to universality and discrimination?

A

poor on both

20
Q

How did psbK-psbI contribute to universality and discrimination?

A

good discrimination poor sequence

21
Q

How did rpoC1/B contribute to universality and discrimination?

A

good universality poor discrimination

22
Q

How do you write the volume dial for a p200? What is its range?

A

xxx|x, 20-200uL

23
Q

How do you write the volume dial for a p1000? What is its range?

A

xxxx 200-1000uL

24
Q

How do you write the volume dial for a p20? What is its range?

A

xx|xx 2-20uL

25
Q

If you have two true breeding parents what will the genotype of the offspring be?

A

heterozygous for both traits

26
Q

How do you determine how much agarose to add based on given volume and percentage?

A

multiply volume by percent
ex. 95mL and 9% gel= 95(.009)=855g

27
Q

According to Wu et al., 2018 what 3 things contribute to hearing loss?

A

exposure to amino glycoside antibiotics, A1555G mutation in the mitochondrial 12S rRNA, T15943C mutation in the mitochondrial tRNA The

28
Q

What does penetrance mean?

A

The ratio of individuals with the genotype, that also display the corresponding phenotype

29
Q

What are three key structural functions of a tRNA molecule?

A

bind to amino acid, bind to ribosome proteins, bind to mRNA codon

30
Q

According to Wu et al., 2018 how many mtDNA mutations did individuals affected with hearing loss have?

A

22

31
Q

What is a cryptogram?

A

A plant that reproduces via spores and not flowers and/or seeds

31
Q

Which two loci were able to amplify over 80% of all land plant samples?

A

rbcL and trn-psbA

32
Q

Did the DNA barcoding loci include plastid and nuclear encoded genes?

A

No

33
Q

Of all the DNA samples that they isolated sequence from, what percentage represents the amount for which they obtained data from all 7 loci?

A

43%

34
Q

What makes for an ideal DNA barcode?

A

routine retrieval with a single primer pair

35
Q

How does the barcoding paper define universality?

A

A locus that can be routinely sequences across most if not all land plants

36
Q

What are 4 reasons to not use the turnH-psbA locus?

A

mononucleotide repeats, manual editing of sequence, lack of bidirectional unambiguous sequences, low quality sequence

37
Q

What are the 5 functions of the m7G cap for mRNA?

A

splicing, polyadenylation, protection from degradation, nuclear export, translation initiation

38
Q

In the RNAi experiment what were the two functions that NAD+-RNA did not have in humans?

A

protection from degradation and translatability

39
Q

According to the RNAi paper is there NAD+-RNA specified localization in plants?

A

Yes, more common in inflorescence (flowers) compared to leaves

40
Q

What are the 3 characteristics of genes that produce NAD-RNAs?

A

fewer introns, small amount are non-coding, shorter in length

41
Q

What are two reasons as to why chloroplasts could not product NAD-RNA?

A

Localized RNA polymerase might not be able to add NAD+ and NADP/NAPH has a higher relative abundance

42
Q

What are the 4 steps of using insert/gene specific primers?

A

Isolating DNA, discuss which primers to use, PCR for confirmation, expected results

43
Q

If there is nothing irregular about the sequence of the insert can you use insert specific primers?

A

Yes you can use all primers, insert specific, gene specific, and insert/gene specific hybrid. Do a traditional genotype experiment

molecular genetics (5 decks)

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