Team:Cambridge/Experiments/Assembly of Reflectin Constructs

This is a placeholder. We should fill it in.

Construct Design

Primer Design

We should mention expected lengths of products here.

Assembly: first attempt

PCR

In the first round of PCR, we amplified fragments required for the assembly of GA1, GA2, GA3 and GA4 constructs.

• We performed PCR using Phusion Hot Start DNA Polymerase in 20 μl reaction volume.
• The time profile used in the PCR machine was the following:

• We decided to use 55 degrees annealing temperaure, although the calculated temperature for most primers is 5-10 degrees higher, because of low annealing temperature of the VF2 primer.

Gibson Assembly

Transformation

Results

Diagnostics

Assembly: second attempt

PCR

Gibson Assembly

Transformation

Results

What next?