| Sample Type/Format |
Sample Requirements |
Remarks |
| Plasmid |
- 80-200 ng/µl
- 10 µl for initial reaction, 3 µl for
each of additional reaction |
DNA size 4-8kb : 80-120 ng/µl
DNA size 10-15kb : 150 - 200 ng/µl |
| PCR Product |
- 5-40 ng/µl
- 10 µl for initial reaction, 3 µl for
each of additional reaction
|
Size 0.5-1 kb : 5-20 ng/µl
Size 1-2 kb : 20-40 ng/µl |
| BAC |
- 500 ng/µl - 1 ug/µl
- 15 µl for initial reaction, 6 µl for
each of additional reaction
|
Size 40-200 kb |
| 16S rDNA sequencing |
- Agar plates or glycerol stock
- genomic DNA: 30-50 ng/µl
- 15 µl of genomic DNA, 200 µl of glycerol stock
|
N/A |
| Primer Walking |
- 8 µl (for an insert size
of up to 4kb) |
Single Strand Sequencing : 1µl
kb insert
If insert size is longer than 4kb,
clone is required in an agar stab
culture status.
|
| Primer |
- 2-5 pmol/µl
(10pmol/µl for BAC/Cosmid samples)
- 10 µl for initial reaction, 2 µl for
each of additional reaction |
18-27 mer, Tm 55°C - 60°C
Universal Primers are available at
free of charge |
| Premix |
| Plasmid + Primer |
- 8 µl (640 ng-1600 ng)
+ 4 µl (8 pmole) |
|
| Purified PCR product + primer |
- 8 µl (40-320 ng) + 4 µl
(8 pmole) |
|
Sequencing Sample/Primer Requirements
