Telephone: 815 753 0638
Cost is $10 per sample for NIU scientists
Fax: 815 753 7855
email: sgrayburn@niu.edu
Name_____________________________________________________ Phone_______________
Email address_______________________________________________ Date _______________
Account to be billed:_________________________________________
Sample
ss/ds/pcr? Size in kb
Primer Name Comments
insert/vector
1 _______________ _________ ____________ ___________ __________________________
2 _______________ _________ ____________ ___________ __________________________
3 _______________ _________ ____________ ___________ __________________________
4 _______________ _________ ____________ ___________ __________________________
5 _______________ _________ ____________ ___________ __________________________
Primers available at the core facility:
T7: 5' TAA TAC GAC TCA CTA TAG GG 3'
SP6: 5' ATT TAG GTG ACA CTA TAG 3'
T3: 5' AAT TAA CCC TCA CTA AAG GG 3'
MF: (M13 -21 forward primer): 5' TGT AAA ACG ACG GCC AGT 3'
MR1: (M13 reverse primer): 5' AAC AGC TAT GAC CAT G 3'
MR2: (M13 reverse primer): 5' TTT CAC ACA GGA AAC AGC 3'
polyT: 5' TTT TTT TTT TTT TTT (ACG)NN
Notes:
-It is the requestor's responsibility to obtain approval for use of
recombinant DNA from the appropriate institutional biosafety committee
-DNA should be clean and of uniform size before sequencing.
-Template that is adequate for manual sequencing may not be pure enough
for automated sequencing.
-If template DNA is purified using an untested protocol, supply a single
sample to determine if modifications are needed.
-Supply double-stranded (ds) plasmid DNA at 250 ng/ uL in water
-Supply PCR products at 30 ng / uL in water. Primers and free dNTPS
must be removed prior to sequencing.
-Supply single-stranded (ss) DNA at 50 ng / uL in water.
-Label each tube on the top.
-Custom sequencing primers should be at least 18 bases long, have melting
temperatures above 45 C, and lack potential secondary structure. Provide
custom primers at 10 ng/ uL in water and include the sequence or quantity
of each base under `Comments'. Assume 1 ODU (A260)=33 ug of
ss oligonucleotide in 1 mL water.