Team:HKJS S/InterLab

HOME

TEAM

Team Members

Collaborations

PROJECT

Description

Design

Experiments

Notebook

InterLab

Model

Results

Demonstrate

Improve

Attributions

PARTS

Parts Overview

Basic Parts

Composite Parts

Part Collection

SAFETY

HUMAN PRACTICES

Human Practices

Education & Engagement

AWARDS

Applied Design

Entrepreneurship

Hardware

Measurement

Model

Plant

Software

JUDGING FORM ⇗

InterLab

Introduction

In order to attain bronze medal requirement, IGEM teams have to participate in the InterLab study. Laboratories from around the world conducted these experiments. In this manner, their studies are trustworthy and repeatable, the key component of scientific discipline. Thus, other related labs will be able to build upon others’ work.

Indeed, fluorescence is a property commonly measured in synthetic biology, but data from different labs are not comparable due to different units have been used. The data has been processed differently and the outcome is given in ‘relative expression’. The InterLab study aims to counter this issue by using detailed protocol and data analysis to yield absolute units for measuring GFP. This year, we are focusing on reducing variable fluorescence measurements between different labs by normalizing to absolute cell count or colony-forming units (CFUs) instead of OD.

Experiments

❏ Instrument brand and model _BMG FLUO star OPTIMA__ ❏ Can your instrument measure both absorbance and fluorescence? _YES____ ❏ Does your instrument have pathlength correction, and if yes can it be disabled? _YES; YES_________________________________________________________ ❏ Does your instrument have variable temperature settings, and if yes can this be set to room temperature? YES; YES ❏ What filters does your instrument have for measuring GFP? You will need information about the bandpass width (530 nm / 30 nm bandpass, 25-30nm width is recommended), excitation (485 nm is recommended) and emission (520-530 nm is recommended) of this filter. 485nm; 520nm ❏ Does your instrument use top or bottom optics (i.e. does your plate reader read samples from the top of the plate or the bottom)? _Can read from both direction

Results

Calibration

OD600 reference point

	LUDOX CL-X	H2O
Replicate 1	0.172	0.124
Replicate 2	0.185	0.106
Replicate 3	0.176	0.137
Replicate 4	0.172	0.124
Arith. Mean	0.176	0.123
Corrected Abs600	0.054
Reference OD600	0.063
OD600/Abs600	1.178

Particle standard curve

Number of Particles	2.35E+08	1.18E+08	5.88E+07	2.94E+07	1.47E+07	7.35E+06	3.68E+06	1.84E+06	9.19E+05	4.60E+05	2.30E+05	0
Replicate 1	1.017	0.603	0.394	0.284	0.226	0.196	0.182	0.172	0.157	0.164	0.164	0.098
Replicate 2	1.012	0.623	0.398	0.289	0.244	0.212	0.19	0.179	0.176	0.173	0.172	0.101
Replicate 3	0.99	0.62	0.399	0.288	0.242	0.206	0.18	0.175	0.173	0.164	0.164	0.107
Replicate 4	1.093	0.57	0.377	0.28	0.23	0.2	0.183	0.186	0.174	0.176	0.168	0.123
Arith. Mean	1.028	0.604	0.392	0.285	0.236	0.204	0.184	0.178	0.170	0.169	0.167	0.107
Arith. Std.Dev.	0.045	0.024	0.010	0.004	0.009	0.007	0.004	0.006	0.009	0.006	0.004	0.011
Arith. Net Mean	0.921	0.497	0.285	0.178	0.128	0.096	0.077	0.071	0.063	0.062	0.060

Fluorescein standard curve

Fluorescein uM	10.00	5	2.5	1.25	0.625	0.313	0.156	0.078	0.039	0.0195	0.0098	0
Replicate 1	2.327E+04	1.229E+04	6.462E+03	3.505E+03	1.802E+03	9.030E+02	4.610E+02	2.390E+02	1.240E+02	6.100E+01	3.100E+01	1.000E+00
Replicate 2	2.402E+04	1.211E+04	6.585E+03	3.380E+03	1.821E+03	8.900E+02	4.860E+02	2.420E+02	1.160E+02	6.800E+01	3.300E+01	1.000E+00
Replicate 3	2.405E+04	1.224E+04	6.671E+03	3.334E+03	1.739E+03	8.830E+02	4.690E+02	2.490E+02	1.230E+02	6.200E+01	3.800E+01	2.000E+00
Replicate 4	2.301E+04	1.231E+04	6.560E+03	3.403E+03	1.709E+03	8.800E+02	4.520E+02	2.380E+02	1.210E+02	6.100E+01	3.000E+01	1.000E+00
Arith. Mean	2.359E+04	1.224E+04	6.570E+03	3.406E+03	1.768E+03	8.890E+02	4.670E+02	2.420E+02	1.210E+02	6.300E+01	3.300E+01	1.250E+00
Arith. Std.Dev.	5.267E+02	9.108E+01	8.600E+01	7.227E+01	5.256E+01	1.023E+01	1.445E+01	4.967E+00	3.559E+00	3.367E+00	3.559E+00	5.000E-01
Arith. Net Mean	2.359E+04	1.224E+04	6.568E+03	3.404E+03	1.767E+03	8.878E+02	4.658E+02	2.408E+02	1.198E+02	6.175E+01	3.175E+01

Cell Measurement

Fluorescence Raw Readings:

Hour 0:	Neg. Control	Pos. Control	Device 1	Device 2	Device 3	Device 4	Device 5	Device 6	LB + Chlor (blank)
Colony 1, Replicate 1	72	82	125	99	70	137	130	84	70
Colony 1, Replicate 2	67	85	124	97	70	134	127	82	73
Colony 1, Replicate 3	69	84	130	102	67	114	124	82	70
Colony 1, Replicate 4	69	84	130	102	67	114	124	82	70
Colony 2, Replicate 1	77	81	118	103	64	141	121	82	68
Colony 2, Replicate 2	67	81	118	102	72	137	121	81	68
Colony 2, Replicate 3	73	83	120	101	76	136	122	85	68
Colony 2, Replicate 4	70	85	121	104	70	123	121	67	73

Hour 6:	Neg. Control	Pos. Control	Device 1	Device 2	Device 3	Device 4	Device 5	Device 6	LB + Chlor (blank)
Colony 1, Replicate 1	83	1370	201	2086	123	260	180	1240	61
Colony 1, Replicate 2	83	1367	191	2166	123	240	178	1211	66
Colony 1, Replicate 3	90	1312	198	2154	120	230	173	1198	63
Colony 1, Replicate 4	88	1057	199	2074	120	193	170	1217	61
Colony 2, Replicate 1	93	1422	138	2231	118	440	160	1191	63
Colony 2, Replicate 2	86	1302	127	2195	123	399	152	1183	62
Colony 2, Replicate 3	85	1336	126	2303	120	414	150	1169	59
Colony 2, Replicate 4	86	1203	121	2195	116	399	150	1169	61

Abs600 Raw Readings:

Hour 0:	Neg. Control	Pos. Control	Device 1	Device 2	Device 3	Device 4	Device 5	Device 6	LB + Chlor (blank)
Colony 1, Replicate 1	0.114	0.106	0.109	0.106	0.109	0.104	0.104	0.103	0.109
Colony 1, Replicate 2	0.121	0.116	0.114	0.114	0.113	0.11	0.112	0.108	0.112
Colony 1, Replicate 3	0.119	0.112	0.115	0.117	0.114	0.112	0.112	0.111	0.112
Colony 1, Replicate 4	0.127	0.121	0.124	0.119	0.124	0.122	0.12	0.134	0.115
Colony 2, Replicate 1	0.121	0.116	0.116	0.116	0.12	0.115	0.116	0.12	0.123
Colony 2, Replicate 2	0.112	0.121	0.12	0.119	0.126	0.115	0.119	0.116	0.118
Colony 2, Replicate 3	0.116	0.113	0.119	0.118	0.117	0.114	0.112	0.113	0.114
Colony 2, Replicate 4	0.123	0.118	0.118	0.12	0.12	0.115	0.118	0.114	0.117

Hour 6:	Neg. Control	Pos. Control	Device 1	Device 2	Device 3	Device 4	Device 5	Device 6	LB + Chlor (blank)
Colony 1, Replicate 1	0.82	0.79	0.144	0.756	0.823	0.122	0.128	0.817	0.11
Colony 1, Replicate 2	0.809	0.835	0.155	0.798	0.829	0.126	0.135	0.809	0.111
Colony 1, Replicate 3	0.84	0.794	0.192	0.784	0.829	0.149	0.134	0.81	0.127
Colony 1, Replicate 4	0.862	0.66	0.155	0.772	0.866	0.121	0.158	0.811	0.153
Colony 2, Replicate 1	0.881	0.83	0.124	0.802	0.834	0.171	0.12	0.832	0.135
Colony 2, Replicate 2	0.863	0.803	0.125	0.822	0.858	0.187	0.128	0.83	0.128
Colony 2, Replicate 3	0.85	0.817	0.139	0.824	0.833	0.187	0.142	0.825	0.142
Colony 2, Replicate 4	0.863	0.739	0.148	0.812	0.835	0.188	0.147	0.825	0.143

Colony-Forming Unit

	p1			p2			n1			n2
D3	_	_	_	282	_	_	_	_	_	_	_	_
D4	124	154	135	91	62	80	116	95	120	116	184	103
D5	21	40	17	12	78	20	13	20	10	12	3	10