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Concentration and Quality Control

This service is only for future sequencing projects at CTG.


RNA


RNA concentration

Fluorometric measurement instruments of nucleic acids are based upon the use of fluorogenic dyes that bind selectively to RNA. This gives a more accurate concentration estimate than spectrophotometric methods as Nanodrop.
At CTG we use Qubit for this application.

RNA quality

Micro-capillary gel electrophoresis based systems are used to analyze the integrity of nucleic acids. A quality score is calculated using software algorithms allowing the classification of total RNA. The score is numbering system from 1 to 10 with 1 being the most degraded and 10 being the most intact.
At CTG we use TapeStation or Bioanalyzer for this application. 

Sample Requirements

To be discussed

Deliverables

Concentration measurement results will be delivered as:
•    Excel file containing concentrations
Quality measurement results will be delivered as:
•    PDF files containing electropherogram and RIN values
•    Excel file containing RIN values

 

DNA


DNA concentration

Fluorometric measurement instruments of nucleic acids are based upon the use of fluorogenic dyes that bind selectively to DNA. This gives a more accurate concentration estimate than spectrophotometric methods as Nanodrop.

At CTG we use Qubit or Fluoroskan Fl plate reader for this application.

Sample Requirements

To be discussed.

DNA Quality qPCR (FFPE samples)

A qPCR is performed with a protocol based on Illuminas TruSeq FFPE DNA library Prep QC Kit performed on Quant Studio 3.
It gives an estimation of fragmentation status and amplification potential through the obtained ∆Cq value. It is a protocol for formalin-fixed paraffin-embedded (FFPE) extracted gDNA.

qPCR sample requirements

  • Sample concentration needs to be based on Qubit measurement (fluorometric method)
  • Samples with concentrations lower than 2.5 ng/µl cannot be processed
  • Sample needs to be normalized in EB-buffer (10 mM Tris-Cl, pH 8.5)
    • Concentration: 2.5 ng/µl
    • Volume: 5 µl
  • Samples should be delivered in 200 µl strips
  • All samples should be labeled individually, directly on side and lid of tube
  • Samples must be delivered on ice

Deliverables

Concentration measurement results will be delivered as:

  • Excel file containing concentrations

Quality measurement results will be delivered as:

  • Excel file containing ∆Cq score and quality estimation.

 

Contact for service

E-mail: CTGservice [at] med [dot] lu [dot] se
 

 

Contact for service

CTGservice [at] med [dot] lu [dot] setarget="_self"

Qubit Flex
Qubit Flex