Difference between revisions of "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!