Team:Cornell/Protocol
From 2011.igem.org
Protocols
- Below, please find the steps that we followed to carry out molecular cloning, create recombinant DNA parts, and construct microfluidic channels.
- Below, please find the steps that we followed to carry out molecular cloning, create recombinant DNA parts, and construct microfluidic channels.
Molecular Cloning Protocols
- PCR Reaction
- Note: Keep everything on ice and add all volumes in a PCR tube.
- 38.5μL ddH2O
- 5.0μL 10x buffer
- 2.5μL dNTPs
- 1.0μL forward primer
- 1.0μL reverse primer
- 1.0μL template
- 1.0μL Vent DNA polymerase
- 50.0μL Total
- Based on primers, set an appropriate annealing temperature
- Note: Keep everything on ice and add all volumes in a PCR tube.
- Agarose Gel Electrophoresis
- Prepare a 1% weight-to-volume agarose gel
- PCR Reaction
Microfluidics Protocols
Biosafety Rules and Procedures
We complied with Weill hall’s safety requirements in gaining access to lab space, as well as in use of the lab. All safety information and procedures are linked on the main Weill safety page [http://blogs.cornell.edu/whfs/weill-hall-safety-links-and-information/ here].
Weill Hall Safety Personnel
Scott D. Emr is the director of Weill Hall which is the building our lab is located. There is also a Weill Hall Safety Committee. While we did not personally meet as a group with either groups, we talked Dr. Archer who is in charge of our particular lab space. She approved of our project and helped us with the safe construction of our microfluidic mask.
Safety Training We received safety training from two online courses that all members were required to pass in order to gain access to the building. These were Lab Safety and Chemical Waste Disposal. Also we received training from our lab instructor on basic safety issues such as waste disposal, use of the fuse hood, etc.