Term
| What is gel electrophoresis used for? |
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Definition
| separation of nucleic acids or proteins |
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Term
| What does separation in gels depend on? |
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Definition
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Term
| When can gel electrophoresis be used in the clinical lab? |
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Definition
| cancer mutations, genetic variations, proteins that cause disease (nonsense mutations) |
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Term
| What are the type types of materials to make gels? |
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Definition
| agarose and polyacrylamide |
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Term
| What is good about agarose gel having large pores? |
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Definition
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Term
| What is bad about agarose gel having large pores? |
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Definition
| bands tend to be fuzzy and spread far apart due to pore size |
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Term
| What can polyacrylamide gels be used for? |
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Definition
| gene sequencing and protein and enzyme analysis |
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Term
| Why do polyacrylamide gels have thin bands? |
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Definition
| gradient gels provide a continuous decrease in pore size from top to bottom |
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Term
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Definition
| electric current from electrodes which attracts and repel molecules |
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Term
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Definition
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Term
| What does the rate of migration depend on? |
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Definition
| strength of the current, size of molecules and gel percentage |
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Term
| Which molecules move fastest and furthest through the pores? |
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Definition
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Term
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Definition
| good separation of large fragments about 5-10kb in size |
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Term
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Definition
| good resolution for small fragments about 0.2-1kb in size |
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Term
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Definition
1. weight out agarose
2. add agarose to 250mL conical flask
3. add 50mL of 1X TAE
4. Microwave at 30 second intervals until clear
5. allow it to cool until comfortable to touch
6. add EtBr
7. Pour the gel slowly into the tank, pushing air bubbles to the side with pipette tip
8. insert the comb, let sit for 30 minutes
9. remove the comb |
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Term
| what do you submerge the gel in? |
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Definition
| 1X TAE bufer, 3-5mm (same buffer you made the gel with) |
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Term
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Definition
| allows for identifying molecular weight, helps identify whats in the sample |
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Term
| what is the direction of electrophoresis? |
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Definition
| negative (black) to positive (red) |
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Term
| when do you stop the gel? |
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Definition
| when it has run 3/4 the length of the gel |
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