|
GSL - Sequencing
Auburn
University Genomics & Sequencing Lab(AU-GSL) uses the ABI
3100 Genetic Analyzer for DNA sequencing. It is required that
DNA submitted for sequencing is already purified by Qiagen
or Millipore, or
Promega's miniprep kit. No
need for plasmid DNA purification IF plasmid DNA has been amplified
with "TempliPhi™
DNA Sequencing Template Amplification Kit". No samples
will be returned. Please include 8 microliter DNA for each reaction.
(see chart below). You can view your results by using "Chromas"
(for windows) or "Vector
NTI " (for Mac). Please send or mail your samples to Room
33, Life Science Building, Auburn University. Auburn, AL 36849.
Sequencing Preparation Chart
DNA
concentration required for PCR products
| PCR
|
100-200 bp |
200-500 bp |
500-1000 bp |
1000-2000 bp |
>2000 bp |
| |
1-3 ng/ul |
3-10 ng/ul |
5-20 ng/ul |
10-40 ng/ul |
40-100 ng/ul |
DNA concentration
required for plasmid
| DNA
Template |
3-5
Kb |
5-8
Kb |
8-10 Kb |
>10
Kb |
|
Double Stranded |
100-200 ng/ul |
250-500 ng/ul |
500-800 ng/ul |
1 ug/ul |
|
Single Stranded |
25-35 ng/ul |
50-70 ng/ul |
100-140 ng/ul |
200 ng/ul |
AU
GSL Primers: AU GSL provides following primers for sequencing
| Primer
|
Sequence
|
| SP6
promoter primer |
5'-GAT
TTA GGT GAC ACT ATA G-3' |
| T3
promoter primer |
5'-ATT
AAC CCT CAC TAA AGG GA-3' |
| T7
promoter primer |
5'-TAA
TAC GAC TCA CTA TAG GG-3' |
|
M13 Reverse primer |
5'-TCA CAC AGG AAA CAG
CTA TGA C-3' |
|
M13 Forward primer |
5'-TGT AAA ACG ACG GCC
AGT-3' |
Customer
Provided Primers: Primer concentration should be 3 uM or 20 ng/ul.
Please provide 8 ul of primer
per reaction
Order
Form
Sequencing
Prices: $10.00/sample
1-5 (96-well) plates: 25% discount ($7.50/sample, or
$720/plate) 6-10 (96-well) plates:30% discount ($7.00/sample, or $672/plate)
11-15 (96-well) plates:35% discount ($6.50/sample, or
$624/plate) 16-20 (96-well) plates:40% discount ($6.00/sample, or $576/plate)
21-25 (96-well) plates:50% discount ($5.00/sample, or
$480/plate) 25-49 (96-well) plates: 60% discount ($4.00/sample, or $384/plate)
50-99 (96-well) plates:65% discount ($3.50/sample, or
$336/plate) 100-199 (96-well) plates: 70% discount ($3.00/sample, or $288/plate)
200+ (96-well) plates: 75% discount
($2.50/sample, or $240/plate) | 
