What should this page have?

• Chronological notes of what your team is doing.
• Brief descriptions of daily important events.
• Pictures of your progress.
• Mention who participated in what task.

Using DNA distribution kit plates: Rehydration of registry DNA from kit plate

Wells in the DNA distribution kit plates contain 2-3 ng of dry DNA. This should be enough DNA for a few transformation, but insufficient for immediate restriction and ligation.

Materials

• DNA distribution kit plates
• Sterile distilled water
• Micropipette
• Sterile micropipette tip

Protocol

1. Locate the well with the DNA part you need
2. Using a micro pipette, draw 10 μL sterile distilled water into a sterile pipette tip
3. Use the pipette tip to puncture a hole through the cover foil and into the well containing DNA you need.
4. Release the water into the well and pipette up and down a few times to rehydrate the DNA you need
5. Store the re-hydrated DNA in the -20C freezer. You can either leave it in the original plate, or transfer it into a separate tube (make sure to label the tube!).
6. The rehydrated DNA can be used immediately to transform competent cells. As little as 1 μL should be enough for one transformation.

Restriction Digest

This protocol is for cutting approximately 500ng of plasmid with two different restriction enzymes. This should provide enough cut DNA for both ligation and agarose gel electrophoresis.

Materials

• Plasmid DNA
• 10x Restriction buffet
• Sterile distilled water
• Restriction Enzymes(EcoR1, Spe1, Xba1, and Pst1)
• Thermocycler
• (0.6 mL) Microcentrufuge tube
• Microcentrifuge
• Ice
• Micropipettes and Sterile pipette tips
• Water bath and 37C