Difficult Template sequencing (SS1006)

This service caters specifically to templates with sequence context difficulties. Sequence context difficulties include high G%, high C%, high GC%, repeat regions and loop structures found in shRNA vectors. 1st BASE adopts a robust protocol that maximizes the success rate of sequencing through these regions. However, the success rates of GT and GC repeats have shown to be limited by this chemistry. Currently, there is no guaranteed to overcome such homopolymeric regions. Please refer our troubleshooting guide for more details.

Specifications

As compared to our regular sequencing services, a greater volume for each sample is to be submitted (in case a repeat run is required).Sample Preparation Requirements for Local Customers (Singapore and Malaysia) are as follows: 

Type of DNA Concentration & Volume/ Reaction
Purified PCR product [90-250bp] 10 ng/µL, min 10 µL per rxn in dH2O
Purified PCR product [251-500bp] 20 ng/µL, min 10 µL per rxn in dH2O
Purified PCR product [>500bp] 40 ng/µL, min 10 µL per rxn in dH2O
Purified Plasmid 100 ng/µL, min 10 µL per rxn in dH2O
Purified M13/phagemid ssDNA 50 ng/µL, min 10 µL per rxn in dH2O
Purified Genomic DNA 400 ng/µL, min 20 µL per rxn in dH2O

 

Type of Primer Concentration & Volume/ Reaction
Primer 10µM or 10pmol/µL, in 10µL per rxn in dH2O

Sample Preparation Requirements for International Customers. 

We recommend that international customers send a larger volume of their samples to overcome loss of samples due to evaporation during the extended transportation to our facilities.

Type of DNA Concentration & Volume/ Reaction
Purified PCR product [90-250bp] 10 ng/µL, min 20 µL per rxn in dH2O
Purified PCR product [251-500bp] 20 ng/µL, min 20 µL per rxn in dH2O
Purified PCR product [>500bp] 40 ng/µL, min 20 µL per rxn in dH2O
Purified Plasmid 100 ng/µL, min 20 µL per rxn in dH2O
Purified M13/phagemid ssDNA 50 ng/µL, min 20 µL per rxn in dH2O
Purified Genomic DNA 400 ng/µL, min 40 µL per rxn in dH2O

 

Type of Primer Concentration & Volume/ Reaction
Primer 10µM or 10pmol/µL, in 10µL per rxn in dH2O