Team:Stony Brook/Notebook/Week9
Week 9
7/22:
• Miniprep of liquid cultures
7/23:
• Restriction Enzyme Digest of miniprepped plasmids and inserts
• Ran all digests on gels (1.2% for vectors and 1.8% for inserts)
• Gel extracted, LnqZE T did not work
• Made more liquid cultures of MBP, SUMO, LnqZE T, and LnqZE U
7/24:
• Nanodrop of gel extracts
• Restriction Enzyme Digest
• Minipreps of liquid cultures
• Restriction enzyme digests
• Ran on gels
7/25:
• Ran on gels
• Finally have enough vector and insert to ligate hybrid inserts into vectors
• Make dilutions of gel extract inserts to perform ligations (Tagged in MBP and Untagged in SUMO)
• Outreach at Stony Brook Movie Night to educate the community about synthetic biology and iGEM and promote our Chipotle fundraiser!
7/26:
• Transformation of DH5α E. coli with MBP-LnqZA and MBP- LnqZE tagged hybrids, and untagged LnqZE and LnqZA.
• Made our lysis, wash, and elution buffers.
• Made small scale (5 mL) liquid cultures of GST lnqZ, GST A53, SUMO Epi, and SUMO LnqZ
• Chipotle fundraiser from 5-9 pm!
7/27:
• Made large scale (1 L) liquid cultures of GST lnqZ, GST A53, SUMO Epi, and SUMO LnqZ
• Induced our large scale liquid cultures with IPTG at mid-logarithmic (OD600 around 0.70) growth stage
• Centrifuged large scale cultures to collect cell pellet. Store at -80°C
• Made small scale liquid cultures of MBP hybrids
7/28:
• Presented about iGEM at Stony Brook’s Science and Society Undergraduate College Freshman Orientation
• Miniprep of MBP hybrids
• Preparation of miniprepped MBP hybrids and sent for sequencing!
