TBL Genomics

Question 1

Dot blot analysis

Answer 1

Simplification of Northern, Southern, Western blots. Molecules not separated by electrophoresis. Mixture containing molecule to be detected applied directly on membrase as a dot. Then detected by probes or antibodies. Saves time, but no information on the size of target molecule–only confirm presence or absence.

Question 2

Restriction fragment length polymorphism (RFLP)

Answer 2

Combination of restriction enzyme digestion of DNA and gel electrophoresis, may use Southern blot

Question 3

Restriction endonucleases

Answer 3

Derived from bacteria and cleave foreign DNA but not native (methylation protects it). Cleavage site specificty with repeats 4-6bp long. Makes sticky ends or blunt ends.

Question 4

Allele Specific Oglionucleotides (ASO)

Answer 4

Short pieces of synthetic oglionucleotide DNA (15-21bp) that are complimentary to target DNA. Used as probe to detect presence of target DNA in Southern blot or dot blot. Specific for only one allele. Must be labeled with radioactive, enzymatic, or fluorescent tag.

Question 5

Variable number tandem repeats (VNTR)

Answer 5

Short repeated DNA sequences. ATTCG repeated more than 3x becomes VNTR. Number of repeats can be used for parental identification. Produce a pattern of bands unique to each individual, but should share similar bands in relatives.

Question 6

cDNA library

Answer 6

Transcripts of structural genes (mRNA) that are expressed by cells. Library can be tissue-specific. mRNA is used to synthesize cDNA with reverse transcriptase.

Question 7

Genomic library

Answer 7

Entire DNA of an organism. Contains sequences for structural genes, introns, regulatory components, noncoding sequences.