Sanger sequencing and genotyping
Fast processing service orders submitted from Feb 22 at 12:00 will be delivered on Feb 25.
Sequencing full service
For ordering full sequencing You'll have to provide us with:
- Template DNA - 300-500 ng of plasmid per reaction, 30-50 ng PCR product per reaction. We can use up to 5,7 μL of template DNA in each reaction mixture
- Primers - for one sequencing reaction we require 0,3 μL of 10 µM (10 pmol/mL) primer (but please send no less than 2-3 μL)
- Complete the order form and send it to kapillaar [ät] ebc.ee; print it out as well and send with DNA and primers to the address
Or bring your samples to the freezer by the security table at our building's Riia street entrance.
We offer following universal primers:
Primer | Sequence | bp |
---|---|---|
ITS4 | TCCTCCGCTTATTGATATGC | 20 |
ITS5 | GGAAGTAAAAGTCGTAACAAGG | 22 |
M13F | GTAAAACGACGGCCAGT | 17 |
M13F-pUC(-40) | GTTTTCCCAGTCACGAC | 17 |
M13R | GCGGATAACAATTTCACACAGG | 22 |
M13R-pUC(-40) | CAGGAAACAGCTATGAC | 17 |
pJET1.2F | CGACTCACTATAGGGAGAGCGGC | 23 |
pJET1.2R | AAGAACATCGATTTTCCATGGCAG | 24 |
SP6 | ATTTAGGTGACACTATAG | 18 |
T3 | ATTAACCCTCACTAAAG | 17 |
T7 | AATACGACTCACTATAG | 17 |
T7promoter | TAATACGACTCACTATAGGG | 20 |
T7terminator | GCTAGTTATTGCTCAGCGG | 19 |
Ready-to-load sequencing
We also analyze ready-to-load sequencing reaction products. Products must be manufactured using the BigDye® Terminator v3.1 (in exceptional cases You can also use BigDye® Terminator v1.1) and diluted in 10 μL formamide (70% formamide or Applied Biosystems Hi-Di formamide). There is no need to fill the order form, please send us the list of sample names in *.txt file, along with invoice recipient.
Fragment analysis
We use a variety of fragment analysis methods - microsatellite analysis, MLPA analysis etc. The client generally prepares the samples to a ready-to-load state, we perform the denaturation before analysis. There is no need to fill the order form, please send us the list of sample names in *.txt file, along with invoice recipient and used dye set.
You can use following dye combinations:
Dye Set |
Filter |
Blue |
Green |
Yellow |
Red |
Size standard |
DS-30 |
D |
6FAM |
HEX |
NED |
ROX |
|
DS-33 |
G5 |
6FAM |
VIC |
NED |
PET |
LIZ |
DS-02 |
E5 |
dR110 |
dR6G |
dTAMRA |
dROX |
LIZ |
Promega |
F |
FL |
JOE |
TMR |
CXR |
|
Custom |
4Dye |
6FAM |
HEX |
TAMRA |
ROX |
|
Custom |
5Dye |
FL |
JOE |
TMR-ET |
CXR-ET |
WEN |
Note that the size standard is added to each sample. For best quality results be sure to use Applied Biosystems Hi-Di Formamide as a loading solution (minimum 10 μL). Minimal order size is 16 samples.
Fast processing service
With fast processing service we guarantee the results by the next working day. Note that the samples must arrive by 9 am if You want to receive results within the same working day. For ordering fast processing service, please write it to the "Comments" section in the order form.