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Sample Requirements

 
 
Buffer Recommendations 
DNA: 10 mM Tris-HCl buffer (pH 7.5 – 8.5)
RNA: 10 mM Tris-HCl buffer (pH 7.0)
Important: Please avoid any EDTA containing buffer for Nextera and Nextera XT Libraries!
 
DNA/RNA Quantification
DNA/RNA quantification is recommended to be done by a fluorometric method, e.g. PicoGreen®, RiboGreen®, Qubit® etc.
 
Sample Amounts Required for Illumina Sequencing
Type of Library Amount (µg) Concentration (ng/µl)
DNA for Nextera XT library >0.2 >10
DNA for TruSeq library >0.5 >10
DNA for Mate Pair library  >3 >30
RNA for RNASeq library (poly(A) enriched) >1 >20
RNA for RNASeq library (ribo depletion) >3 >20
DNA for amplicon preparation (e.g. 16s rDNA or ITS analysis) >0.1 >05
1st or 2nd step Nextera PCR product >0.2 >05
Ready-to-sequence amplicon library * >0.2 >10
ChIP-Seq libraries (req. ChIP DNA) >0.05 >05
miRNA/small RNA (req. total RNA) >3 >200
low input RNA (req. total RNA) >0.01 >01
 
*)  for single samples to be pooled > 50 ng > 5 ng/µl per sample
 

Please ship DNA/RNA samples in plate format for more than 24 samples, otherwise we have to charge additional costs. If you plan to provide indexed libraries, please make sure to use Illumina compatible index combinations at an early stage of your project.

Sample naming: Please use short, unique names without special characters. If bioinformatics analysis is desired we use the names as is for the provided tables and figures.

Sample Amounts Required for PacBio Sequencing
Type of Library Amount (µg) Concentration (ng/µl)
DNA Library >10 >300
 

with an OD260/OD280 ratio of 1.8 to 2.0. High molecular weight DNA and pure samples are critical for library preparation and a good data quality.

Therefore please:

  • Avoid multiple freeze-thaw cycles
  • Do not expose it to high temperatures
  • Do not use DNA intercalating stains such as ethidium bromide and/or UV light  since they can induce DNA damage
  • Avoid containing insoluble material.
  • Avoid containing RNA contamination.
  • Avoid containing denaturants (e.g. guanidinium salts or phenol) or detergents (e.g. SDS or Triton-X100).
  • Avoid incubation in complex or rich media
  • Harvest during early- to mid-log-phase, from several rather than a single culture
 
DNA Quality Control
Microsynth performs on each received sample a complete quality control prior to further sequencing
steps. However, we do recommend that you also check your DNA on a gel or on the Bioanalyzer.
If you do so, please provide us with the gel picture or trace file as well.
 
Sample & Data Storage at Microsynth
Data, Samples and the processed libraries will be stored at Microsynth for 6 months.