Week 9: Monday June 27 - Sunday July 3

Monday

We created a set of lab rules due to common mistakes made in the lab over the past few weeks.

Vicki and Marianne will having training sessions throughout the week to show team members without much wetlab experience how to PCR, cast and run a gel, digest, ligate, transform, design primers, send miniprepped products to be sequenced.

Modeling: Jacob obtained the ARC program. Gurpal wants to create a wetlab flow chart to match the modeling.

Sent DspB (part from last year) to Grinnell College (a new iGEM team this year who requested our part) in the United States.

alpha-pinene synthase

• SDM-PCR worked! (Used the QuikChange SDM Kit protocol).

3-carene synthase

Daisy needs to make custom primers for sequencing purposes because the mutagen site is in the middle of the coding sequence and her flanking primers will only sequence 700-800bp into the sequence.

beta-pinene synthase

• Resuspended SDM forward and reverse primers

(-)-limonene synthase

• Resuspended SDM forward and reverse primers

Tuesday

Training: went over PCR protocol (SDM specific)

beta-pinene & (-)-limonene synthase

• Ran SDM-PCR

3-Carene

Daisy did a single digest with EcoRI-HF to test if the EcoRI site is in the synthase or not. Chris Keeling emailed Daisy the sequence he used for the truncation of the 3-Carene synthase. Daisy ordered primers to do the sequencing. However, Alina has not replied to her email to confirm that the primers are ordered.

Wednesday

Training:

Thursday

Training:

Friday

Saturday

Sunday