Barnes - gels etc.

Question 1

What are the main types of gel, and the subtypes of these?

Answer 1

• agarose or polyacrylamide

- native or denaturing

Question 2

What are diff % agarose gels used for?

Answer 2

• high % for smaller things (better sep)

- low % for bigger things

Question 3

What type of mols are agarose gels used for?

Answer 3

• DNA/RNA

Question 4

What type of gel is a sequencing gel?

Answer 4

• denaturing polyacrylamide gel

Question 5

What are native or denaturing gels used for?

Answer 5

• native = looking at complexes

- denaturing = info about prot size

Question 6

What kind of substrate might the mol looking at be?

Answer 6

• DNA/RNA/prot etc.

Question 7

Where might the substrate come from?

Answer 7

• gDNA/oligo/cell lysate from bacteria etc.

Question 8

How could the substrate be detected?

Answer 8

• radioactive label/coomassie/silver etc.

Question 9

When would a probe be used?

Answer 9

• if looking for a specific thing

Question 10

What diff steps should you go through when answering a Q?

Answer 10

1) what kind of gel?
2) what kind of substrate and where does it come from?
3) how is substrate being detected?
4) diff between the diff reactions (controls)?
5) how do these diffs affect any products formed?

Question 11

What is the diff between endo and exonucleases?

Answer 11

• exonucleases cleave nts 1 at a time from end of polypep chain
• endonucleases cleaves phosphodiester bonds in middle of polypep chain