User:KCliff/Evaluating Cross Contamination Using The Pin Tool

In preparation for the 2008 iGEM competition, selected DNA 'bioparts' found in the Registry are transferred onto sheets of thesis paper using the pin tool replicator in order to be mailed to teams at participating colleges and universities. Since this is the first year that iGEM will be using the thesis paper in place of welled plates to distribute the 'bioparts', the pin tool replicator method of transfer was evaluated for possible DNA and culture cross contamination.

Cross Contamination Among DNA Samples from the Same Source Plate

Materials:

• 96-deep well plate
• 25% Cresol Red
• 96 Multi-Blot (Pin Tool) Replicator
• source plate Library Copier
• Thesis Paper with iGEM 96-block grid
• Lint-free blotting paper
• Pin Cleaning Solution
• 99% Isopropynol

1. Add 1.0 ml of 25% Cresol Red solution to the wells of every other column on the deep well plate.

2. Transfer the 25% Cresol Red solution to the thesis paper using the Pin Tool Replicator.

3. Clean the pin tool replicator.

4. Observe spotting on thesis paper grid to determine cross contamination among boxes.

Culture Cross Contamination Among Samples from the Same Source Plate

Materials:

              96-deep well plate (2)
              96 Multi-Blot (Pin Tool) Replicator
              LB broth
              Red Florescence Protein (RFP) cells
              Library Copier (2)
              Lint-free blotting paper 
              Pin Cleaning Solution
              99% Isopropynol

1. Add 500 ul of LB broth to each well in a 96-deep well source plate.

2. To every other column of wells add 10 ul of suspended RFP cells, leaving the alternate columns with only LB broth.

3. Using the Pin Tool Replicator, Transfer the culture from the source plate to a transfer 96-deep well plate of LB broth.