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Journal 12.09.10:. Which sizes of DNA fragments do you expect to get from the digestion? 2. How can one use the electrophoresis procedure to estimate the size of the DNA fragments?. pARA-R construct. Recombinant plasmid of interest. pARA-R 4720 bp. pARA-R. 4720 bp. Bam H I. rfp 702bp.

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Which sizes of dna fragments do you expect to get from the digestion

Journal 12.09.10:

  • Which sizes of DNA fragments do you expect to get from the digestion?

  • 2. How can one use the electrophoresis procedure to estimate the size of the DNA fragments?


Which sizes of dna fragments do you expect to get from the digestion

pARA-R construct

Recombinant plasmid of interest

pARA-R

4720 bp

pARA-R

4720 bp

BamH I

rfp

702bp

rfp 702bp

Hind III


Which sizes of dna fragments do you expect to get from the digestion

Gel Electrophoresis - Detection

We run the gel until the yellow dye runs out.

Wells


Which sizes of dna fragments do you expect to get from the digestion

After running the gel (till the yellow dye runs out), we stain it in Ethidium bromide, which will fluoresce when bound to DNA:


Which sizes of dna fragments do you expect to get from the digestion

Supercoiling stain it in Ethidium bromide, which will fluoresce when bound to DNA:

Multimers

“nicked-circle”

circle


Which sizes of dna fragments do you expect to get from the digestion

Structural forms of plasmid stain it in Ethidium bromide, which will fluoresce when bound to DNA:

Slower than nicked.

“multimer”

“nicked-circle”

Linear

(Digested)

Supercoiled


Which sizes of dna fragments do you expect to get from the digestion

A- stain it in Ethidium bromide, which will fluoresce when bound to DNA:

A+

10 Kb Ladder

10 Kb Ladder

10 Kb Ladder

Multimer

Nicked

Super Coiled

5 Kb

Linear Fragment

Linear Fragment


Which sizes of dna fragments do you expect to get from the digestion

M stain it in Ethidium bromide, which will fluoresce when bound to DNA:

A-

A+

Yellow highlighter shows group number..

Expected results


Which sizes of dna fragments do you expect to get from the digestion

Class Results 2013: Lab 4a stain it in Ethidium bromide, which will fluoresce when bound to DNA:


Which sizes of dna fragments do you expect to get from the digestion

Groups 1,2 stain it in Ethidium bromide, which will fluoresce when bound to DNA:


Which sizes of dna fragments do you expect to get from the digestion

Groups 3,4 stain it in Ethidium bromide, which will fluoresce when bound to DNA:


Which sizes of dna fragments do you expect to get from the digestion

4000base pairs stain it in Ethidium bromide, which will fluoresce when bound to DNA:

2000base pairs

1500 base pairs

1000 base pairs

500 base pairs


Which sizes of dna fragments do you expect to get from the digestion