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   & Fragment Analysis
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B065 and B017
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Next Generation Sequencing
 

SAMPLE SUBMISSION REQUIREMENTS

Before submitting your order online, please follow our minimum requirements for the samples being submitted for each service below:


Full Service: (back to top)

• Sample QC --
You need to check the quality of your samples before submitting them to us. QC can be done on Agilent Bioanalyzer, and the data can be submitted through our online order form.

• Required Amount of Starting Material --

The amount required will be dependent of the library preparation kit chosen. PAN is currently supporting sample prep kits from Illumina.If you are unsure of which kit to use, we will work with you to determine the kit best suited for your project.

The following table can be used as a guide to choose an appropriate kit for your project.
(Click on the name of each kit for more detailed information)

Starting Material
Application
Project Type
Sample Prep Kit Option(s)
Sample Type
Input Quantity
• DNA
DNA Sequencing
Whole Genome
*TruSeq DNA (LT or HT) gDNA 1ug
*TruSeq DNA PCR-Free (LT or HT) gDNA 1-2ug
Nextera DNA gDNA 50ng
Nextera Mater Pair gDNA 1-4ug

** Targeted Resequencing
(Enrichment)
-Hybrid capture
-Metagenomics
(16S rRNA)
-Clone checking

Nextera Rapid Capture Exome gDNA 50ng
Nextera Rapid Capture Expanded Exome gDNA 50ng
TruSeq Custom Enrichment gDNA 1ug
Nextera Custom Enrichment gDNA 50ng
Nextera Rapid Capture Custom Enrichment gDNA 50ng
Small Genome
Nextera XT gDNA 1ng
Amplicon
Nextera XT PCR amplicons (at least 300bp) 1ng
TSCA gDNA 250ng
TSACP gDNA,
FFPE DNA
150-250ng

 

• Gene Regulation Analysis


 

ChIP-SEQ

 

 

TruSeq ChIP -Seq

 

 

ChIP DNA

 

 

5-10ng

 

• RNA
• Transcriptome Analysis
mRNA-SEQ
TruSeq RNA v2 Total RNA 1-4ug
mRNA entire fraction of mRNA
*TruSeq Stranded mRNA (LT or HT) Total RNA 0.1-10ug
mRNA 10-400ng
Total RNA-SEQ
( mRNA + non-coding RNA)
*TruSeq Stranded Total RNA (LT or HT) Total RNA
(human, mouse or rat or other eukaryote)
0.1-1ug
Low quality or
FFPE RNA
0.1-1ug
Small RNA
TruSeq Small RNA (48rxns) small RNA entire fraction of small RNA purified from 1-10ug of total RNA.

*TruSeq LT: 24-plex; TruSeq HT: 96-plex
** For exome capture, you need to specify the total number of samples for pre-enrichment pooling.

• Library QC on MiSeq --

We offer further QC of the prepared library(ies) on our MiSeq machine. For more information, you can refer to the information in the MiSeq Run section below.



MiSeq Run : (back to top)

Sample QC --

It is important that you check the quality and quantity of your samples before submitting them to us. For best results and accurate readings, use Bioanalyzer and Qubit to determine the library's size and concentration, respectively. We will ask for the results if they are availble.

We will also run your sample on our Agilent Bioanalyzer as part of our initial QC step for MiSeq Run. If the sample failed QC and/or the results are not the same as those provided by you, we may ask for sample resubmission or a written confirmation (email will suffice) stating that it is okay for us to proceed with MiSeq run as is.


Desired Run Cycle and Expected Read Length --

Depending on the application and the read length needed, you need to choose the right run cycle for your sample.

The table below will give you guidance to choose the right run cycle based on the application needed.

Application/Project Type Recommended Read Length
Targeted Resequencing
Amplicon (tens of targets)
Amplicon (hundreds of targets)
Hybrid Capture (thousands of targets)
16S Metagenomics
Clone Checking

1 x 250
2 x 250
2 x 75
2 x 150
1 x 36
Small Genome
De novo
Resequencing
Plasmids

2 x 250
2 x 250
2 x 250
RNA Sequencing
Small RNA
RNA-Seq (microbial)

2 x 25
2 x 50-250
ChIP Seq
2 x 25
Library QC
2 x 25
** For more information on other applications and recommended sample prep kits, please refer to this brochure and Illumina website.

Order Submission --
For the library being submitted, please place an order online, then email us detailed information including library's size, library's concentration, index barcodes and sequences (if any). You can download this template and then email it to Elizabeth Zuo .