{"id":33,"date":"2014-11-08T00:33:56","date_gmt":"2014-11-08T00:33:56","guid":{"rendered":"http:\/\/www.hawaii.edu\/microbiology\/asgpb\/?page_id=33"},"modified":"2025-07-01T09:43:14","modified_gmt":"2025-07-01T09:43:14","slug":"sample-submission","status":"publish","type":"page","link":"https:\/\/www.hawaii.edu\/microbiology\/asgpb\/?page_id=33","title":{"rendered":""},"content":{"rendered":"

\"SAMPLEHEADER\"<\/a><\/p>\n

DNA Requirements for Sanger Sequencing<\/strong><\/p>\n

Mix your template (5 \u03bcl) and primer (1 \u03bcl of 3.2 \u03bcM) according to the specifications below.<\/p>\n\n\n\n\n\n\n\n
DNA Type<\/strong><\/strong><\/p>\n
\n<\/td>\n		Template Length<\/strong><\/strong><\/p>\n
\n<\/td>\n		Template Amount<\/strong><\/strong><\/p>\n
\n<\/td>\n		Primer Amount<\/strong><\/strong><\/p>\n
\n<\/td>\n		Premixed Volume<\/strong><\/strong><\/p>\n
\n<\/td>\n<\/tr>\n
Plasmid (vector + insert)<\/td>\n1 kb \u2013 10 kb<\/td>\n200 ng<\/td>\n3.2 pmol<\/td>\n\u2265 6 \u03bcl\/reaction<\/td>\n<\/tr>\n
10 kb \u2013 20 kb<\/td>\n400 ng<\/td>\n<\/tr>\n
PCR products & DNA fragments<\/td>\n100 \u2013 500 bp<\/td>\n5 \u2013 20 ng<\/td>\n<\/tr>\n
500 \u2013 1000 bp<\/td>\n20 \u2013 50 ng<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n
* For large DNA such as plasmids over 20 kb, Lambda, Cosmids, and BACs, please contact us for further details before submitting for sequencing.<\/address>\n
\u00a0<\/address>\n
\u00a0<\/em><\/strong>Note:<\/em><\/strong><\/address>\n
If you want to submit your template and primer in separate tubes, provide 10 \u03bcl of template and 10 \u03bcl of primer (diluted to 3.2 \u03bcM). This is double the amount of template and primer volume that we actually need so that we can rerun samples if there are any problems. Your templates and primers will be kept for one week in case you need them returned back to you.<\/p>\n
\n<\/address>\n

\u00a0Supplied Primers<\/strong><\/p>\n

ASGPB provides the primers listed below at no additional cost:<\/p>\n\n\n\n\n\n\n\n\n\n
Primer<\/strong><\/span><\/td>\nAbbreviation<\/strong><\/span><\/td>\nSequence (5′ to 3′)<\/strong><\/span><\/td>\n<\/tr>\n
M13 Forward<\/td>\nM13F(-20)<\/td>\nGTA AAA CGA CGG CCA GT<\/td>\n<\/tr>\n
M13 Reverse<\/td>\nM13R(-27)<\/td>\nCAG GAA ACA GCT ATG AC<\/td>\n<\/tr>\n
T7 Promoter<\/td>\nT7<\/td>\nTAA TAC GAC TCA CTA TAG GG<\/td>\n<\/tr>\n
T7 Terminator<\/td>\nT7 ter<\/td>\nGCT AGT TAT TGC TCA GCG G<\/td>\n<\/tr>\n
T3<\/td>\nT3<\/td>\nAAT TAA CCC TCA CTA AAG GG<\/td>\n<\/tr>\n
SP6<\/td>\nSp6<\/td>\nATT TAG GTG ACA CTA TAG<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n

The following are some tips to help design your custom primers<\/strong>:<\/p>\n

The primer should be at least 15 to 20 bases from the insert<\/p>\n

The primer should be between 18 to 25 bases long<\/p>\n

Aim for ~50% GC content, with the Gs & Cs evenly distributed throughout the primer<\/p>\n

Avoid runs of identical nucleotides<\/p>\n

The Tm should be 55\u00b0C to 75\u00b0C<\/p>\n

Use software to check your primer for any dimers or hair-pin loops that may be formed<\/p>\n

\n

\u00a0<\/strong>Sample Submission<\/strong><\/p>\n

Fill out the Sequencing Order form and submit with your samples to:<\/p>\n

ASGPB<\/address>\n
University of Hawai\u2018i at M\u0101noa<\/address>\n
1800 East-West Road, Life Sciences Building 201<\/address>\n
Honolulu, HI 96822<\/address>\n

<\/h2>\n

Drop off samples Monday through Friday between 9:30 a.m. to 5:30 p.m.<\/p>\n

\u00a0\u00a0\u00a0 \u00a0\u00a0 \u00a0 *If you are shipping samples to us, use the following mailing address:<\/p>\n

\u00a0\u00a0 \u00a0 \u00a0 \u00a0 ASGPB<\/address>\n
\u00a0\u00a0\u00a0 \u00a0 \u00a0\u00a0 University of Hawai\u2018i at M\u0101noa<\/address>\n
\u00a0\u00a0\u00a0 \u00a0 \u00a0\u00a0 3190 Maile Way, St. John 101<\/address>\n
\u00a0\u00a0 \u00a0 \u00a0 \u00a0 Honolulu, HI 96822<\/address>\n
\u00a0<\/address>\n
\n

Data Delivery<\/strong><\/p>\n

You will be notified by e-mail with your finished sequences.<\/p>\n

You should receive the results in one or two business days unless there are unforeseen delays. In that event, you will be notified as soon as possible after the samples arrive at the facility.<\/p>\n

The results for each sequence will be provided in abi format files. The ABI file (.abi) contains the nucleotide sequence, electropherogram, and other information about the run conditions. This file requires software such as Chromas, 4Peaks, Geneious, etc to view it.<\/p>\n

\n

Contact<\/strong><\/p>\n

If you have any questions or would like more information about our other services, please contact Shaobin Hou or Jennifer Saito.<\/p>\n

Email: asgpb07@gmail.com or asgpb@hawaii.edu<\/p>\n

Phone: (808) 956-8039<\/p>\n","protected":false},"excerpt":{"rendered":"

DNA Requirements for Sanger Sequencing Mix your template (5 \u03bcl) and primer (1 \u03bcl of 3.2 \u03bcM) according to the specifications below. DNA Type Template Length Template Amount Primer Amount Premixed Volume Plasmid (vector + insert) 1 kb \u2013 10 kb 200 ng 3.2 pmol \u2265 6 \u03bcl\/reaction 10 kb\u2026<\/p>\n

Continue reading<\/span><\/i><\/a> <\/p>\n","protected":false},"author":2,"featured_media":0,"parent":0,"menu_order":0,"comment_status":"closed","ping_status":"closed","template":"templates\/template-onecolumn.php","meta":{"footnotes":""},"class_list":["post-33","page","type-page","status-publish","hentry"],"_links":{"self":[{"href":"https:\/\/www.hawaii.edu\/microbiology\/asgpb\/index.php?rest_route=\/wp\/v2\/pages\/33","targetHints":{"allow":["GET"]}}],"collection":[{"href":"https:\/\/www.hawaii.edu\/microbiology\/asgpb\/index.php?rest_route=\/wp\/v2\/pages"}],"about":[{"href":"https:\/\/www.hawaii.edu\/microbiology\/asgpb\/index.php?rest_route=\/wp\/v2\/types\/page"}],"author":[{"embeddable":true,"href":"https:\/\/www.hawaii.edu\/microbiology\/asgpb\/index.php?rest_route=\/wp\/v2\/users\/2"}],"replies":[{"embeddable":true,"href":"https:\/\/www.hawaii.edu\/microbiology\/asgpb\/index.php?rest_route=%2Fwp%2Fv2%2Fcomments&post=33"}],"version-history":[{"count":55,"href":"https:\/\/www.hawaii.edu\/microbiology\/asgpb\/index.php?rest_route=\/wp\/v2\/pages\/33\/revisions"}],"predecessor-version":[{"id":678,"href":"https:\/\/www.hawaii.edu\/microbiology\/asgpb\/index.php?rest_route=\/wp\/v2\/pages\/33\/revisions\/678"}],"wp:attachment":[{"href":"https:\/\/www.hawaii.edu\/microbiology\/asgpb\/index.php?rest_route=%2Fwp%2Fv2%2Fmedia&parent=33"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}