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Sequencing Services

  
 

Sequence Reaction Guidelines

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1/16th Big Dye Reaction in 5ul
0.5ul of Big Dye Mix
0.75ul of Applied Biosystems 5x dilution buffer (provided with kits)
3pmols of primer
template per the following guidelines
q.s to 5ul

 

Template 

Quantity per Reaction

100–200 bp PCR product

1–3 ng

200–500 bp PCR product

3–10 ng

500–1000 bp PCR product

5–20 ng

1000–2000bp PCR product 

10–40 ng 

>2000 bp PCR product 

20–50 ng

Single-stranded plasmid:

25–50 ng

Double-stranded plasmid:

150–300 ng

Cosmid, BAC

0.5–1.0 µg

Bacterial genomic DNA 

2–3 µg

Cycling Conditions

For 1 cycle:
96° 1min
For 25 cycles:
96° 10sec
50° 5sec
60° 4min
For 1 cycle:
4°   ¥

*Reactions should be submitted  in PCR strip tubes with strip caps. Tubes with individual or attached caps will not be accepted. We recommend strip tubes from Island Scientific, P/N IS-435.

 

 

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