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Reference List of DNA sequencing and Services
Genomic Department
A. DNA sequencing of PCR products# and plasmids up to 750 bases:
|
Nber of Bases |
Specification |
Reference |
|
350 Bases |
Single strand basecalls non corrected |
SSBN 0350 |
|
|
350 Bases |
Single strand basecalls corrected |
SSBC 0350 |
|
350 Bases |
Double Strand |
SDBR 0350 |
|
600 Bases+50 |
Single strand basecalls non corrected |
SSBN 0600 |
|
600 Bases+50 |
Single strand basecalls corrected |
SSBC 0600 |
|
600 Bases |
Double Strand |
SDBR 0600 |
|
750 Bases+100 |
Single strand basecalls non corrected |
SSBN 0750 |
|
750 Bases+100 |
Single strand basecalls corrected |
SSBC 0750 |
|
750 Bases |
Double Strand |
SDBR 0750 |
Universal primers (M13, M13R, T3, T7, T7 terminator, Sp6, CMV forward, BGH Reverse, pGEX3’ , pGEX5’…)
are provided free of charge. For specific primers send your primer(s) (10µM) or the primer sequence(s)
(SSOL 0003).
# purified single amplification product.
Set up:If no data are obtained, the processing cost of 8.00 euros per sequence will be charge (SECH 0001).
Turnaround time for this service is 24 hours
Call us for a special discount according to the quantity
B. Primer walking :
|
Nber of bases |
Reference |
|
Single strand |
1-10 kb |
SMSB 0010 |
|
Double strand for publication (accuracy: 99.99%) (1) |
1-20 kb |
SMDB 0020 |
|
Double strand FDA quality (accuracy: 100%) (1, 2) |
1-20 kb |
SMDB 0030 |
|
Cosmides, BACs, PACs |
> 20 kb |
SMCO 0030 |
1 Complete editing of delivered data, design and synthesis of specific primers, alignment and assembly
of the sequences.
2 FDA quality : Sequencing will be performed to, at least, a 4-fold redundancy, resulting in the required
accuracy level of 100%
C. BAC end and Cosmid extremities sequencing :
|
Specification |
Material |
Reference |
|
Single strand, 300-500 bases (Phred 20). |
DNA (1µg/seq) |
SBAE 0001 |
|
Bac DNA preparation, 2 reads, one of each side, 300-500 bases (Phred 20). |
Clone (glycerol stock or stab) |
SBAE 0002 |
Set up: If no data are obtained, the processing cost of 25% of the total price will be charge (SECH 0003).
D. High throughput sequencing§:
1. High throughput PCR products sequencing §:
|
Nber of Bases |
Specification |
Material |
Reference |
|
700 Bases |
PCR product purified #, single strand, raw data |
Purified PCR product |
SHTN 0650 |
|
700 Bases |
Purification of the PCR mix #, single strand, raw data |
PCR product |
SHPP 0650 |
# Single amplification product checked on agarose gel.
Read lengths of PCR products are dependant of the PCR products lengths
§ High througput sequencing project when order ³ 96 sequences (1 microplates of 96 wells).
§ HTS set up: For sequence < 200 PHRED 20 bases, the processing cost of 5.50 euros per sequence will be charge
(SECH 0002).
2. High throughput Plasmids (EST, SAGE, …) sequencing §:
|
Nber of Bases |
Specification |
Material |
Reference |
|
700 Bases |
Single strand, raw data, average (except empty clones) of 550 readable bases (Phred 20); no minimum |
Plasmid DNA |
SHTP 0650 |
|
700 Bases |
Plasmid DNA preparation, single strand, raw data, average (except empty clones) of 550 readable bases (Phred 20); no minimum |
Clone (glycerol stock) |
SPHT 0650 |
§ High througput sequencing project when order ³ 96 sequences (1 microplates of 96 wells).
§ HTS set up: For sequence < 200 PHRED 20 bases, the processing cost of 5.50 euros per sequence will be charge
(SECH 0002).
3. High throughput Bac end sequencing §:
|
Specification |
Material |
Reference |
|
Single strand, raw data, average (except empty clones) of 400-600 readable bases (Phred 20); no minimum |
DNA (1µg/seq) in 96 plate |
SHBE 0001 |
|
Bac DNA preparation, 1 reads, raw data, average (except empty clones) of 400-600 readable bases (Phred 20); no minimum
Bac DNA preparation, 2 reads, one of each side, raw data, average (except empty clones) of 400-600 readable bases (Phred 20); no minimum
|
Clone (glycerol stock) in 96 plate
Clone (glycerol stock) in 96 plate
|
SHBE 0002
SHBE 0003
|
§ High througput sequencing project when order ³ 96 sequences (1 microplates of 96 wells).
§ HTS set up: For sequence < 200 PHRED 20 bases, the processing cost of 5.50 euros per sequence will be charge
(SECH 0002).
E. DNA purification :
|
Material |
References |
|
Plasmid DNA Mini preparation : 5-10 µg* |
Glycerol stock or fresh culture on LB agar |
SADN 0001 |
|
Plasmid DNA Midi preparation: 50-100 µg * |
Glycerol stock or fresh culture on LB agar |
SMID 0001 |
|
Plasmid DNA Maxi preparation: 250-500 µg * |
Glycerol stock or fresh culture on LB agar |
SMAX 0001 |
|
Plasmid DNA Mega preparation: 1-2.5 mg * |
Glycerol stock or fresh culture on LB agar |
SMEG 0001 |
|
Plasmid DNA Giga preparation: 5-10mg * |
Glycerol stock or fresh culture on LB agar |
SGIG 0001 |
|
Endotoxin-free Plasmid DNA Mega prep: 1-2.5mg * |
Glycerol stock or fresh culture on LB agar |
SENM 0001 |
|
Endotoxin-free Plasmid DNA Giga prep: 5-10mg * |
Glycerol stock or fresh culture on LB agar |
SENG 0001 |
|
DNA preparation (plasmid) in microplate (96 wells)* |
Glycerol stock or fresh culture on LB agar |
SADN 0006 # |
|
Genomic DNA extraction from plant
and fungi
|
Glycerol stock or stab
|
SADN 0008
|
|
BAC DNA preparation for Bac end sequencing* |
Glycerol stock or stab |
SBAC 0001 |
|
Ultrapure BAC DNA preparation (for shot gun bank) |
Glycerol stock or stab |
SBAC 0002 |
Purification of PCR products (single band) |
PCR products (30µl) |
SPCR 0002 |
Agarose gel purification of PCR products (GeneClean) |
PCR products (30µl) |
SPCR 0005 |
|
Agarose gel purification of 0.5µg of DNA insert (GeneClean) |
20µl of 0.2µg/µl of plasmid DNA |
SADN 0008 |
Purification of PCR reaction in plate 96 wells (single band) |
PCR products in plate 96 wells |
SPCR 0004 |
|
RNA extraction |
Frozen tissue |
SRNA 0001 |
|
mRNA purification |
Total RNA |
SRNA 0002 |
|
Template re-purification |
|
SADN 0004 |
Set up : If no data are obtained, the processing cost of 25% of the total price will be charge (SECH 0003).
#Template preparation (plasmid) in microplate (96 wells) drop to 1.50 euros / sample if order is associated with
HTS project.
*Actual yields depend on plasmid copy number, size of insert and host strain
F. Genomic project :
|
|
Material |
References |
|
Synthesis of a primer for sequencing |
Sequence |
SSOL 0003 |
|
Design and synthesis of a primer for sequencing |
|
SSOL 0002 |
|
Single strand sequence up to 600 bases (minimum of 250 bases phred 20) of difficult sample (high % of GC, repeats, hairpins, …) q |
30 µl of 0.2 µg/µl of DNA |
SADN 0007 |
|
Restriction digestion analysis with 1 enzyme (plasmid DNA) ¤ |
20 µl of 0.2 µg/µl of DNA |
SADN 0008 |
|
Restriction digestion analysis with 1 enzyme (plasmid DNA) |
10µg of DNA |
SDIG 0001 |
|
BAC mapping |
|
SCAR 0001 |
|
PCR reaction |
1 µg of DNA in 10µl of H2O (Minimum) |
SPCR 0001 |
|
PCR reaction with a proof reading polymerase |
1 µg of DNA in 10µl of H2O (Minimum) |
SPCR 0006 |
Retro-Transcription |
RNA pellet in alcohol (-70°) |
SRTR 0001 |
Plasmid DNA transformation |
20 ng of Plasmid DNA (20 ng/µl) |
SADN 0005 |
|
Mono-cistronic plasmid |
|
SPLA 0001 |
|
Bi-cistronic plasmid |
|
SPLA 0002 |
|
Tri-cistronic plasmid |
|
SPLA 0003 |
|
Bibliography study for gene isolation and cloning strategy |
|
SCRE 0001 |
|
Design and synthesis of primers for cDNA library screening |
|
SCRE 0002 |
|
cDNA library screening |
|
SCRE 0003 |
|
cDNA library screening (customer library) |
cDNA library |
SCRE 0004 |
|
Protein expression analysis SDS PAGE |
|
SPRO 0001 |
|
Protein expression analysis SDS PAGE - Western Blot* |
|
SPRO 0002 |
PCR product cloned into bacterial system |
Purified PCR product |
SPCR 0003 |
DNA subcloning into bacterial system |
(Call) |
SADN 0002 |
DNA subcloning into yeast system |
(Call) |
SADN 0003 |
¤ Restriction digestion analysis to check if a clone is positive for sequencing.
q Genomic project set up: if no data are obtained, the processing cost of 25% of the total price will be charge
(SECH 0003).
* Antibody against classic Tag (HIS, GST, c-myc, HA). Exotic antibody has to be provided by the customer.
Sequencing Options :
|
References |
|
Colour print of the chromatogram (Price per sequence) |
SDIV 0001 |
|
Information on diskette |
SDIV 0002 |
|
Information on CD-ROM |
SDIV 0003 |
|
Express delivery |
SDIV 0006 |
|
Express delivery in dry ice |
SDIV 0007 |
G. Bioinformatics :
|
Description |
References |
|
Sequence alignment (Price per sequence) |
Alignment of the sequence chromatograms to edit the concensus sequence |
SDIV 0005 |
|
HTS sequence quality analysis (price per plate)
(order ³ 1 microplate 96 wells) |
Phred 20 basecalls, Crossmatch analysis,
insert size (% < 500bp), polyA representation (EST projects) |
BIOA0005 |
| AutoBlast Analysis (price per plate) |
Phred 20 basecalls, Crossmatch (contamination)
Databank formatting
Output results in flat format (text tabulated),
Summary of the results per project and per plate |
BIOA0009
|
|
HTS alignment to the NCBI databases (BLAST)
(Price per plate)
(order ³ 1 microplate 96 wells) |
Phred 20 basecalls, Crossmatch (contamination)
Output results in flat format (text or XML)
Summary of the results per plate (text tabulated format) |
BIOA0010
|
|
HTS alignment to the NCBI databases (BLAST) (Price per plate)
(order ³ 1 microplate 96 wells) |
Phred 20 basecall, Crossmatch (contamination)
Output results in flat format (text or XML),
Summary of the results per project and per plate,
Friendly HTML presentation with links to NCBI (Genbank, LocusLink) |
BIOA0011 |
|
HTS analysis to LocusLink databases (official nomenclature, aliases, sequence accessions, phenotypes, MIM numbers, UniGene clusters, homology, map locations,
Gene Ontology …) |
Output results in table (Excel or HTML) with links to related databases and related web sites.
Price varies according to number of links |
BIOA0020 |
|
Assembly shotgun projects |
Only PHRED 20 basecalls are used
Contamination, Repeat mask, EST clustering
PHRAP or Sequencher is used for assembly of the shotgun sequences |
BIOA0030 |
|
SAGE data analysis |
Extract, count and calculate relative abundance of the tags .
Identification of the tag according to available SAGEmap.
Satistical analysis of the results |
BIOA0040 |
|
Peptide Design |
In collaboration with peptide synthesis department |
BIOA0050 |
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MilleGen Biotechnologies - Genomic Department Price list. Status : September 26, 2003. (Prices do not include delivery costs, taxes and may change without notice).
For ordering please use the ordering forms on our web site: www.millegen.com.
MilleGen, Prologue Biotech, rue Pierre et Marie Curie, BP28262, 31682 Labège (France)
Tel :+ 33 (0)5 61 28 70 10 ; FAX : 33 (0) 5 61 28 70 11; Email : sequence@millegen.com |
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