Bioplatforms Australia Datasets Initiative
IlluminaHiSeq 2000 Sample submission form
ResearcherContact Details
Name:
DrCaroline Moffat
Email:
Caroline.Moffat@curtin.edu.au
Institution / Organisation:
Australian Centre for Necrotrophic Fungal Pathogens (ACNFP)
Sample Information
Organism / Species
Sample type
DNA/RNA
Part of organism RNA/RNA extracted from
Extraction method
Comments / special considerations
Address:
Department of Environment and Agriculture
Curtin University of Technology
Bentley, WA 6102
Fungus (Pyrenophoratritici-repentis), Plant (wheat)
RNA
Mycelia (fungus), Uninfected and Ptr Infected Leaves (plant)
Trizol reagent and column purification; Trizol reagent andcolumn purification followed by LiCl reprecipitation
-
Growth protocol of fungus and/or plant (medium, soil, water regimen, light/day, fertilisers etc):
Fungus - Ptr cultures grown on V8PDA agar; Plant - Wheat seedlings grown on vermiculite without fertiliser
BPA Project: Ramaciotti Sequencing submission form
Phone:
(08)9266 9916 (office)
(08)9266 1876 (lab)
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Treatment protocol (i.e. route of administration of pathogen):
Plant infection assay - whole plant (foliar)spray
Further Information on experimental design (i.e. timepoints and biological replicates):
Fungal mycelia – vegetative and sporulating mycelia harvested at one time point
Plant infection assay – fungal infected and uninfected wheat leaves harvested at three and four DPI, RNA pooled from biological replicates
BPA Project: Ramaciotti Sequencing submission form
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Sample Name
Volume
(ul)
RIN (RNA)
OD
260/280
OD
260/230
Conc.(ng/ul)
(method used)
(NanoDrop
Spectrophotometer)
Sample 1
C.Moffat M1
28
-
2.11
1.98
668.5
Sample 2
C.Moffat M2(a)
16
-
2.05
2.12
414.4
Sample 3
C.Moffat D1
20
-
2.15
1.96
813.0
Sample 4
C.Moffat D2(a)
15
-
2.12
2.42
361.2
Sample 5
C.Moffat C1
23
-
2.12
1.91
550.1
Sample 6
C.Moffat P1
23
-
2.13
2.07
368.0
Sample 7
C.Moffat C2
20
-
2.11
2.22
784.5
Sample 8
C.Moffat P2
20
-
2.09
2.04
314.7
Additional Information.
Vegetative mycelia (isolate Meck4),
7-day-old V8PDA agar plate culture
Sporulating mycelia (isolate Meck4),
9-day-old V8PDA agar plate culture
Vegetative mycelia (isolate DW5),
7-day-old V8PDA agar plate culture
Sporulating mycelia (isolate DW5),
10-day-old V8PDA agar plate culture
Control leaves (3 DPI),
RNA pooled from 3 replicates
Ptr infected leaves (3 DPI),
RNA pooled from 3 replicates
Control leaves (4 DPI),
RNA pooled from 6 replicates
Ptr infected leaves (4 DPI),
RNA pooled from 6 replicates
Attach additional sheet if more samples
Sample Requirements
RNA
•
•
Samples should be intact and not degraded as assessed by a Bioanalyzer. The RNA Integrity Number (RIN) value should be greater than 8.
OD 260/280 ratio of 2, and a 260/230 of 1.8-2.
BPA Project: Ramaciotti Sequencing submission form
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•
5 μg of total RNA at min concentration of 200ng/ul (optimal 500ng/ul). The concentration should be measured using the Ribogreen
fluorescent assay.
Samples should be resuspended in nuclease-free water or elution buffer.
RNA that has been extracted using Trizol or any phenol based method must undergo an additional column purification
•
•
Sample shipment details

Samples to be shipped on dry ice.
Facility
The Ramaciotti Centre
Address
Lowy Cancer Research Centre
C25
via Gate 11 Botany Street
University of New South Wales
Randwick, NSW 2052
Contact person
Tonia Russell
Phone: (02) 93851658
Email: illumina@unsw.edu.au
Please email a copy of the completed form to Anna Fitzgerald (afitzgerald@bioplatforms.com) at the time of sample submission and complete Google Docs metadata form.
BPA Project: Ramaciotti Sequencing submission form
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