NANOPORE SEQUENCING

WHAT WE SEQUENCE:

PLASMIDS, LARGE AMPLICONS, PHAGE, AND BACMIDS. WE ACCEPT COLONY PICKS TOO! EASY!

PLEASE TALK TO US BEFORE YOU SUBMIT SAMPLES FOR NATIVE BARCODING AND LIGATION SEQUENCING.

LIBRARY KITS WE USE / OFFER:

RAPID BARCODING KIT: Tagmentation based barcoding

NATIVE BARCODING KIT: Ligation based barcoding for those require control over read length. PLEASE call or email before you submit samples!

LIGATION SEQUENCING KIT: This kit performs adapter ligation only, with no barcording step. It is designed for samples with know identifiable sequences and where control over read length is required. Input is 1ug of gDNA or 100-20 fmol of shorter fragment. PLEASE call or email before you submit samples!

sample submission deadlines

2PM EVERY TUES - FRIDAY FOR PURIFIED PLASMIDS, AMPLICONS, PHAGE, AND BACMIDS

10AM EVERY TUES - FRIDAY FOR COLONY PICKS AND SAMPLES THAT REQUIRE RCA

DNA quantification guideline:

WE NO LONGER REQUIRE DNA QUANTIFICATION PRIOR TO NANOPORE SEQUENCING!

HOWEVER -- IF YOU HAVE THE ABILITY TO ASSESS THE DNA CONCENTRATION (AND QUALITY), IT IS RECOMMENDED TO PROVIDE THAT INFORMATION ALONG WITH THE SAMPLE (ON THE REQUEST FORM).

IF DOING SO, PLEASE USE A QUBIT ASSAY OR THE PROMEGA QUANTIFLUOR dsDNA SYSTEM TO MEASURE YOUR DNA CONCENTRATION (NO NANODROP MEASUREMENTS PLEASE) - however, see below to download a file for approximating Nanodrop to Qubit concentration numbers.

service rates:

On-Campus and Other UC Campuses $15/sample.

LBNL and CHORI (+10%) $16.50/sample.

Off-Campus (+60.5%) $24.08/sample

Price includes pre-treatment of plasmids, library prep, Nanopore run, and basic bioinformatics

SAMPLE SUBMISSION GUIDELINES:

All samples should be in water or TE:

How much to submit for Rapid and Native Barcording:

Colony picks ~10ul

Purified plasmids 20ul at 50ng/ul or more

Amplicons 20ul at 50ng/ul or more

Bacmids 20ul at 100ng/ul or more

Please email your plasmid sequences in FASTA format for reference mapping if available.

Sample names:

Letters, Numbers, and "_" only.

NO SPACES (e.g., " ") or other characters.

Please use short names, if at all possible.

HOW TO PLACE ORDERS:

· Submit 20ul of >50ng/ul plasmids and amplicons in 1.5 ml Eppendorf tubes or a 96well plate

· Please label the top of the tube with a permanent marker as clearly as possible with alpha-numeric characters only

· Bring your samples to 310 Barker Hall or ship via FedEx

· Fill out and email Full Plasmid Sequencing order form or Full Plasmid Sequencing-Plate order form to dnaseq@berkeley.edu. · · · Please include a copy along with your samples

TURNAROUND TIme:

Runs start every Tuesday through Friday at 2PM.

The results are ready next afternoon, typically by 2PM.

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