Molecular Biology Service Centre
The Molecular Biology Service Centre (MBSC) provides molecular biology support for life science research community in a time- and cost-effective manner. It builds on the idea that time and money can often be saved by consolidating routine or challenging cloning projects into a central facility operated by experienced researchers.
Due to the similarity and routine nature of many DNA cloning projects, they can be efficiently performed in the MBSC on a mid-throughput scale using common reagents and operations, thus reducing time and materials. Using this service rather than doing these tasks in the research lab can thus save time and money, and allow researchers to focus more on the discovery aspects of their research.
Outsourcing routine, specialize operations is not a new concept. For example,
almost everyone uses specialized services for DNA synthesis and sequencing.
Nonetheless, DNA cloning projects are not always routine and it is likely that
graduate students should master essential cloning techniques before depending on
the service.
[+] Services (click on left [+] symbol to expand)
CLONING AND MUTAGENESIS
PCR Cloning : PCR cloning of a DNA sequence in to a specific vector. This can be done even if there is no feasible restriction site to use.
Sub-Cloning : Sub-cloning a DNA sequence into a specific vector. No identified restriction site is required.
Site Directed Mutagenesis (substitution, insertion or deletions of any size): The mutagenesis can be applied to anywhere in a gene or vector. The substitution, insertion or deletion sequence can be any size up to a few kilobases (kb).
Tagging,: Add tags before or after a gene in a construct.
Domain Swapping: Generate chimeric proteins by domain swapping
Vector Construction: custom vector construction.
TRANSFORMATION: Transformation of plasmids into bacteria
PLASMID DNA PREPARATION
Mini prep: users can supply 2ml of culture, OD
600>0.5, colony plates or glycerol stock.
Max prep: users can supply 250 ml of culture, OD
600>0.5, colony plates or glycerol stock.
OTHER MOLECULAR SERVICES PLEASE INQUIRE.
[+] Types of Cloning/Mutagenesis processes (click on left [+] symbol to expand)
Cloning Projects: (Type C Series)
Type C1 cloning includes: (most sub-cloning or PCR projects fall
in this category)
General sub-cloning or PCR cloning into the multiple cloning sites with feasible restriction site(s) to use and only involves the 20 common restriction endonucleases listed below.
(List of the 20 common
restriction endonucleases: BamHI, BglII, ClaI, EcoRI, EcoRV, Hind III, KpnI,
MluI, NcoI, NdeI, PstI, PvuII, SmaI, SacI, SalI, StuI, ScaI, StyI, XbaI, XhoI).
T/A Cloning.
PCR cloning into a unspecified vector (the MBSC vector is a
ampicillin resistant bacteria vector, it can be used for sequencing)
Type C2 cloning includes: (most site directed mutagenesis projects
fall in this category)
General sub-cloning or PCR cloning into the mulitple cloning sites with feasible restriction site(s) to use and involves:
blunt cloning
into a specific vector or
restriction endonucleases BEYOND the 20 common ones listed
listed below.
(List of the 20 common
restriction endonucleases: BamHI, BglII, ClaI, EcoRI, EcoRV, Hind III, KpnI,
MluI, NcoI, NdeI, PstI, PvuII, SmaI, SacI, SalI, StuI, ScaI, StyI, XbaI, XhoI).
Type C3 cloning includes:
General sub-cloning or PCR cloning into the multiple cloning sites but WITHOUT feasible restriction site(s) to use
Type C4 cloning includes:
General sub-cloning or PCR cloning into ANYWHERE in a construct and WITHOUT feasible restriction site(s) to use.
Insert or substitute a sequence into a construct or vector without a restriction site including large mutations. (DNA template needed for the mutated to sequence)
Mutagenesis Projects: (Type M Series)
Type M2 cloning includes:
Point mutation: Site directed mutagenesis with any combination of mutations within a
24 base frame.
Type M3 cloning includes:
Large point mutation: Site directed mutagenesis with any combination of
mutations within a 25 to 48 base frame.
Mutagenesis to make deletions of any size.
Type M4 cloning includes:
General sub-cloning or PCR cloning into ANYWHERE in a construct and WITHOUT feasible restriction site(s) to use.
Insert or substitute a sequence into a construct or vector without a restriction site including large mutations. (DNA template needed for the mutated to sequence)
[+] Fee Schedule and Turnaround Time (click on left [+] symbol to expand)
SERVICE FEES FOR CLONING AND MUTAGENESIS
All cloning material costs are included in the service charge. The fee is
based on cloning types and number of cloning processes.
One cloning process is defined as one of the followings: sub-cloning, PCR cloning, site directed mutagenesis (point mutation, insertion, deletion or substitution). If a construct requires two processes, for example, doing mutagenesis of a gene and then transferring it into a different vector, it is charged as two processes. Fee arrangements for more complex projects can be negotiated, usually on a per hour basis.
Base Fee Schedule (per process): for cloning and mutagenesis
Cloning Type
(see above for
details) |
MBB |
SFU |
Academic |
| Type C1: most TA cloning, sub-cloning, PCR cloning projects |
$105 |
$115 |
$125 |
| Type C2: blunt cloning specific,
sub-cloning or PCR cloning with rare enzymes |
$135 |
$145 |
$160 |
| Type C3: sub-cloning or PCR cloning no
sites to use in MCS |
$180 |
$195 |
$210 |
| Type C4: insertion or substitution
anywhere |
$210 |
$230 |
$250 |
Mutagenesis Type
(see above for
details) |
MBB |
SFU |
Academic |
| Type M2: point mutagenesis:
=24 bp |
$125 |
$135 |
$140 |
| Additional mutation in same construct |
$60 |
$60 |
$60 |
| Type M3: mutagenesis
25-48 bp, deletion mutagenesis any size |
$165 |
$180 |
$190 |
| Type M4: mutation, insertion or substitution
anywhere |
$195 |
$220 |
$240 |
* Base charged is for process with insert or substitute up to 3 kb, no
limit on deletions, and end construct up to 12 kb.
Oversize and Other Charges:
| Cloning Type |
|
| Inset oversize* /extra kb (per extra kb over 3 kb) |
$20 |
| Construct
oversize**/extra kb (per extra kb over 12 kb) |
$20 |
| Antibiotic selection
other than Ampicillin |
$10 |
| GC rich templates |
$20 |
| Additional mutation in
same construct (=24 bp) |
$60 |
| Amplifying from genomic DNA as
template |
$20 |
*Gene Size: the length of the gene in resulting construct, including any tags.
**Construct Size: the size of resulting construct, including vector and gene.
Turn Around Time
First process is around 10-15 working days, i.e. about 2-3 calendar weeks, with primer and
sequencing transit time included; each additional process in same project add
around 5 working days (about 1 calendar week). Note: turn around time is estimate, most projects will
fall within the standard turn around time, but difficult projects may take
longer.
SERVICE FEES FOR OTHER SERVICES
All material costs are included in the service charge. Fee arrangements for more complex projects can be negotiated, usually on a per hour basis.
Fee schedule for other services:
| Service Type |
MBB |
Academic |
| Plasmid mini prep: from supplied grown
culture |
$2.00 |
$2.25 |
| Plasmid mini prep: from colony or
glycerol stocks |
$2.50 |
$2.75 |
| Plasmid maxi Prep: |
inquire |
inquire |
| Other molecular biology services |
inquire |
inquire |
[+] Sequence
Verification and Other Possible Fees (click on left [+] symbol to expand)
Sequence verification: High fidelity polymerase is used for cloning with
an error rate of 1 in 1.3 million base pairs or less. ~750 bp around mutagenesis
site is verified for mutagenesis. There are two
choices of verification of the sequences: self sequence verification and
guaranteed sequence verification.
Self sequence verification: For self sequence verification, you do
the sequence verification of the rest region. However, if you found a
mutation, we will replace a new clone, the request need to be made within 4 weeks upon
delivery of the previous clone.
Guaranteed sequence verification: For guaranteed sequence verification, the
sequence of the insert is guaranteed to match the template you provided, the
charge is $25/extra kb.
DNA templates: Users are usually required to provide starting DNA templates
Other Charges May Occur:
Low quality templates: Plasmids or PCR products can be accepted as
templates. Plasmids need to be purified using a plasmid kit, has a minimal
concentration of 50 ng/ul and a minimal amount of 2 ug. PCR products need
to be clearly one band without other minor bands, a minimal concentration of 25 ng/ul
and minimal amount of 1 ug.
Templates do not meet these standard usually can be accepted with a $20 process surcharge.
Non-standard vectors: Full sequence of the vector or construct shall
be provided; if not, often a vector map with sequences around the multiple
cloning site may work. Otherwise, it may subject to a $40 sites verification
charge for cloning and sequencing.
[+] On Site Supply Centre (click on left [+] symbol to expand)
The following products will be supplied by Molecular Biology Service Centre
The service center also makes the following products at a significant discount compared to commercial products
Pre-made LB agar plates with antibiotics: (10 plates per sleeve): $15/
sleeve
50X TAE gel running buffer. (400ml): $35.
SOC medium for transformation (10ml): $12
Competent bacteria cells for subcloning:
[+] Sample Submission (click on left [+] symbol to expand)
Sample requirements:
Plasmid: (plasmid mini prep kit purified, dissolved
in H2O or Tris buffer:10 mM Tris-HCl pH 8.0) 40 ul with minimum concentration 50
ng/ul,
PCR product (PCR purification kit purified, dissolved in H2O or Tris
buffer:10 mM Tris-HCl pH 8.0) 40 ul with minimum concentration 25 ng/ul
Vector
and PCR products: full sequence required by sending email, if not, then as much
as information as possible.
Templates with a lower concentration maybe
accepted.
Other information required:
Please also send an email with a brief description of the project
including the sequences in a word file and/or plasmid map of all DNA templates as attachments.
Contact
Name: Ziwei Ding
Location: South Science Building 6120, SFU
Phone: 778-782-4090
e-mail: Ziwei Ding zding@sfu.ca
or
Name: Stuart Zong
Phone : 604-818-0169
e-mail:Stuart Zong zzong@sfu.ca
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