Team:MIT/Recombinase Group Notebook Week 3

• BACKGROUND
• EXPERIMENTS
  Synthetic Promoters microRNA Recombinase Proof of Concept Collaborations
• FUTURE WORK
  Circuit Design Clinical Applications
• HUMAN PRACTICES
  Parts Toolbox Accessible Protocols Integrated Human Practices Public Outreach Interlab Study
• NOTEBOOK
  Lab Notebook Parts Mammalian Parts Collection Attributions Safety
• AWARDS
  HP Silver HP Gold Integrated HP Basic Part
• TEAM

JG 6/21/16 midiprep

Made with Benchling

Project: iGem 2016 _ Recombinase subgroup

Authors: Maya Kaul

Date: 2016-06-20

Monday, 6/20

	A	B
1		Concentration
2	EGSH: eYFP	11.7 ng/uL
3	hEF1a: BFP	610.2 ng/uL
4	hEF1a: VgEcr	816.0 ng/uL
5	hEF1a: rtTA	133.4 ng/uL
6	TRE: BFP	765.6 ng/uL

Table1

Midiprep

Introduction

Purifying large amount of DNA to tranfect mammalian cells

Materials

• Qiagen MidiPrep

Procedure

• Overnight Culture 50mL (06/20/16)

1. Prepare the overnight culture 1 day before the midiprep

2. Choose 1 sample for each plasmid constructs from the diagnostic digest result

3. For each sample, preapre a 500mL breathable conical tube

• Add 50mL media

• 5uL of antibiotic

• 10uL of culture from the miniprep

4. Incubate at 37C for 16-20 hours

• Midiprep preparation (06/21/16)

5. Transfer 50mL overnight culture into 50mL Falcon tube

6. Centrifuge at max speed for 15 mins at 4C

7. Following the Qiagen midiprep protocol