Sample Preparation

General Sample Requirements

Sample Type

Volume

Minimum Concentration

Amount

A260/280 Ratio

A260/230 Ratio

RNA Integrity Number

gDNA

>40 µl

20 ng/µl

>2 µg

>1.8

->2

-

Low Input DNA library (such as FFPE)

>40 µl

30 ng/µl

>2 µg

>1.8

>2

-

PCR-free Genomic DNA library

>40 µl

20 ng/µl

>2 µg

>1.8

>2

-

Total RNA

>40 µl

50 ng/µl

>3 µg

>1.8

>2

> 7.5

Small RNA/microRNA library

>40 µl

50 ng/µl

>3 µg

>1.8

>2

> 7.5

It is preferable to quantify the DNA and RNA quantities using fluorometric methods, including Qubit and PicoGreen.  If a spectrophotometer (e,g. Nanodrop) is used, we suggest submitting twice the requested amount of samples due to the unreliable detection of this measurement method.  

gDNA samples

  1. Submit DNA sample(s) in Distilled water or Dissolved in 10mM Tris HCL, pH7.5-8.5 at in a clearly labelled 1.5/2.0ml microcentrifuge tube tightly sealed and parafilm wrapped.
  2. Can be delivered in an ambient temperature or with ice pack.

Total RNA samples

  1. Submit RNA samples(s) in DEPC treated water, or Dissolved in 10mM Tris HCL, pH7.5-8.5 in a clearly labelled 1.5-2.0ml microcentrifuge tube tightly sealed and parafilm wrapped.

    Note: For small RNA analysis, total RNA must be prepared using the appropriate method to ensure that the small RNA fraction is not removed during the purification process. E.g. many column-based procedures have a cut-off size that will eliminate all small RNA, and are not suitable for this purpose.
     
  2. Dry ice shipping: We recommend using a minimum of 8-10kg of dry ice which should last for 24-48hours.

Pre-made Library pools Specification*:

*Note: Please only submit pre-made libraries after discussing with our team.  

Requirements:

Libraries should be submitted in clearly labelled 1.5 microcentrifuge tubes, tightly sealed with parafilm.  Please use low-retention tubes if you have them. The preferred buffer for sequencing libraries is 10mM Tris, 0.1% Tween 20 or 10mM Tris, pH=8.0-8.4. Water can be used as well.

Although we will run the sample on TapeStation 4200 before loading the samples onto the sequencer, please evaluate library concentration, size and quality on your side and provide us with the QC result.

Insert size of libraries (>300bp) : >3ug, Index/MID information to be provided in order form if applicable.