Sequencing

Company

Genewiz:

http://www.genewiz.com/

Username/password: seq@excellgen.com/1234qwer

for <6kb construct, mix  10 ul of 50~100 ug/ul DNA with 2.5 ul 10 uM sequencing primer, +2.5 ul H2O

Sequencing primers are in the boxes labelled with Sequencing Primers in -20 oC freezer

http://www.genewiz.com/Public/Resources/Sample-Submission-Guidelines/Sanger-Sequencing-Sample-Submission-Guidelines/Sample-Preparation#sanger-sequence

Place reaction mix tubes in a plastic bag, stabled onto the order form and place in a blood box (west end near loading dock outside the building)

Macrogen

http://www.macrogenusa.com/

excellgen/1234qwer

Plasmid

Plasmid must be free from salt and ethanol. Qiagen or Omega kit should be used for making plasmid.

Premix:

DNA conc: 10 ng/ul/kb. For a 6 kb plasmid the plasmid DNA concentration should be 60 ng/ul.

pET vector: at least 25 ug/ul. 5kb--> 50 ug/ul.

Adenovirus: 5 kb: 50 ug/ul.

Lentivirus: 50 ~90 ug/ul.

AAV: ~40 ug/ul

10ug/ul/kb: 5 kb template, at least 5 ul, 50 ug/ul.

Use our primer: add 1.3 ul of 10 uM primer to 5 ul of  template, total: at least 6.5 ul