Bone samples Flashcards

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1
Q

What is one way to prepare a bone sample?

A

freezer milling (cryogenic)

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2
Q

Many extraction processes begin with steps aimed at?

A

removing contamination

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3
Q

After decontamination, the hard tissues are incubated in extraction buffer and?

A

proteinase K

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4
Q

What are some decontamination processes for bone samples?

A
  • physically removing the outer bone surface by immersing in a bleach solution
  • exposure to UV radiation
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5
Q

Why are the bone samples grinded to a powder?

A

increases the surface area to the various chemicals in DNA extraction to get more DNA

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6
Q

What is widely used for extraction of DNA from bone samples?

A

phenol-chloroform

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7
Q

After cleaning the bones with abrasion what is the next step to clean them?

A

soak in detergent

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8
Q

In the ball milling extraction process of bone samples what is used to freeze the sample?

A

liquid nitrogen

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9
Q

What are the two type of DNA recovery is used with bone samples?

A
  1. mortar and pestle
  2. cryogenic
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10
Q

What are the benefits of the cyrogenic method over the mortar and pestle?

A
  • preservation of DNA integrity
  • efficient cell distribution
  • reduced contamination
  • consistency in particle size
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11
Q

What are the advantages of the mortar and pestle method over cyrogenic?

A
  • cost effective
  • simple and traditional
  • suitable for small samples
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12
Q

Why would we want to decalcify bone samples?

A
  • facilitate cell lysis
  • prevent DNA degradation
  • improve efficiency
  • enhance purity
  • prevent interference
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13
Q

What are the overall steps in recovering DNA from bone samples?

A
  1. sample prep - clean exterior and remove contaminants
  2. decalcification using EDTA to remove calcium ions
  3. distruption of bone tissue - cryogenic/mortor and pestle
  4. cell lysis
  5. protein digestion - proteinase K
  6. DNA extraction using phenol-chloroform
  7. DNA purification
  8. quantification using a spectrophotometer or fluorometer
  9. quality check - agarose gel or capillary electrophoresis
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