DNA Lab
“When finally interpreted, the genetic messages encoded within our DNA molecules will provide the ultimate answers to the chemical underpinnings of human existence” James D. Watson.
The CEINGE DNA LAB technology platform provides multiple tools to "read" these genetic messages and to "write" new ones thanks to its two sectors Sanger Nucleic Acid Sequencing Service and Oligonucleotide Synthesis Service.
SANGER SEQUENCING SERVICE
The Sanger CEINGE Sequencing Service, thanks to its long experience in automated Sanger sequencing processes, offers maximum flexibility for your samples that require customized solutions in tubes and plates.
We guarantee the interpretation of repeated sequences, whether they are short tandem repetitions (STR or microsatellites) or short and long interval repeats (SINE and LINE), as well as structural rearrangements.
The analytical quality guaranteed for the single reading is 99% accuracy for the first 700 bases for single sequencing and one error every 10000 bases for sequencing projects.
Our primary purpose is to provide our users, regardless of the objective difficulties presented by the specific sample or sequencing project, our support to give them the best possible result.
- TUBE SEQUENCING SERVICE
-Premix Tube Sequencing Service: DNA samples resuspended in water, plasmids and PCR, premixed with specific primers and organized in 1.5 ml Eppendorf tubes (maximum of eight) or in strips of eight tubes.
The template must consist of purified DNA in one of the following concentrations:
-Plasmid-DNA up to 10 kb: 100 ng / μl, final volume of 8 μl PLUS 8 μl of the specific Primer at a concentration of 2 μM. Ensure that the total volume of the DNA is at least 16 μl. The DNA sample must be quantized to the NanoDrop.
-Plasmid-DNA over 10 kb: 100 ng / μl, final volume of 16 μl PLUS 8 μl of the specific Primer at a concentration of 2 μM. Ensure that the total volume of the DNA / Primer mixture is 24 μl. The DNA sample must be quantized to the NanoDrop.
- Purified PCR products: 3 ng per 100 bp in a final volume of 20 μl PLUS 8 μl of the specific primer at a concentration of 2 μM. Make sure that the total volume of the DNA / Primer mixture is 28 μl. The DNA sample must be quantized to the NanoDrop. 
- ORDER MUST BE ACCOMPANIED BY A MOD SS_01 FORM (PROVIDED BY THE FACILITY STAFF) DEBTLY COMPLETED OR PRINTED BY THE REQUEST FOR EXPLANATORY GOALS.
- Mark each tube with the sample number corresponding to the one entered in the MOD SS_01 form (provided by the facility staff) or with the request number, using a permanent marker.
- Mark the first tube of the eight test tubes using an indelible marker and arrange the samples in the same way as indicated in the MOD SS_02 form (provided by the facility staff).
- In case the samples are > 30kbp and/or contain sections that are difficult to read, please report it in the MOD SS_01 form (provided by the facility staff). Special PCR conditions will be adopted in order to obtain the best possible sequencing results.
- Have samples sent to be sequenced at room temperature in our laboratories.
- The results will be sent by e-mail within the second day following the receipt of the order.
- A free repetition is foreseen in case of technical failure.
- Tube sequencing service: provides sequences of DNA samples resuspended in water, plasmids and PCR, using standard service primers and organized in 1.5 ml Eppendorf tubes (maximum of eight) or in eight-tube strips.
The template must consist of DNA at one of the following concentrations:
- Plasmid-DNA up to 15 kb: 100 ng / μl, final volume of 20 μl.
- PCR non-purified products: 100-200 bp: 5 ng / μl; 300-400 bp: 10 ng / μl; 500-600 bp: 16 ng / μl; > 1000bp 20ng / μl final volume of 20 μl.
- Purified PCR products: 3 ng per 100 bp in a final volume of 20. The DNA sample must be quantized to the NanoDrop.
- ORDER MUST BE ACCOMPANIED BY A MOD SS_01 FORM (PROVIDED BY THE FACILITY STAFF) DEBTLY COMPLETED OR PRINTED BY THE REQUEST FOR EXPLANATORY GOALS.
- Mark each tube with the corresponding sample number entered in the MOD SS_01 form (provided by the facility staff) or with the request number, using a permanent marker.
- Mark the first tube of the eight test tubes using an indelible marker and arrange the samples in the same way as indicated in the MOD SS_02 module (to be requested from the facility staff).
- Quantize the templates to be purified by agarose gel or QIAXEL, using a suitable MOLECULAR WEIGHT AND CONCENTRATION WEIGHT marker. The image produced by QIAxcel, on the other hand, must contain a maximum of thirteen samples, so as to allow clear interpretation and must report the default date entered by the instrumentation.
- In case the samples are > 30kbp and / or contain sections that are difficult to read, please report it in the MOD SS_01 form (provided by the facility staff). Special PCR conditions will be adopted in order to obtain the best possible sequencing results.
- Have samples sent to be sequenced at room temperature in our laboratories.
- The results will be sent by e-mail within the second day following the receipt of the order.
- A free repetition is foreseen in case of technical failure.
- DNA mold and sorted primers are stored for two weeks.
- PCR purification is included in the price.
• PLATE SEQUENCING SERVICE
- Premix Plate Sequencing Service: DNA samples resuspended in water, plasmids and PCR, premixed with specific primers and organized in plate.
The template must consist of purified DNA in one of the following concentrations:
-Plasmid-DNA up to 10 kb: 100 ng / μl, final volume of 8 μl PLUS 8 μl of the specific Primer at a concentration of 2 μM. Ensure that the total volume of the DNA / primer mixture is at least 16 μl. The DNA sample must be quantized to the NanoDrop.
-Plasmid-DNA over 10 kb: 100 ng / μl, final volume of 16 μl PLUS 8 μl of the specific Primer with a concentration of 2 μM. Make sure that the total volume of the DNA / primer mixture is 24 μl. The DNA sample must be quantized to the NanoDrop.
- Purified PCR products: 3 ng per 100 bp final volume of 20 μl PLUS 8 μl of your customized Primer with a concentration of 2 μM. Ensure that the total volume of the DNA / primer mixture is 28 μl. The DNA sample must be quantized to the NanoDrop.
- ORDER MUST BE ACCOMPANIED BY A MOD SS_01 FORM (PROVIDED BY THE FACILITY STAFF) DEBTLY COMPLETED OR PRINTED BY THE REQUEST FOR EXPLANATORY GOALS.
- Please arrange the samples in the plate in the same way indicated in the MOD SS_02 form (provided by the facility staff).
- In case the samples are > 30kbp and / or contain sections that are difficult to read, please report it in the MOD SS_01 form (provided by the facility staff). Special PCR conditions will be adopted in order to obtain the best possible sequencing results.
- The results will be sent by e-mail within the second day following the receipt of the order.
- A free repetition is foreseen in case of technical failure.
- The concentration must be measured by NanoDrop and normalized through the plate.
- The plates can contain different types of DNA samples arranged in columns from the first to the twelfth and from the top to the bottom.
- The size of PCR products should not vary by more than a factor of 3.
- The H12 position of the plate must be kept free for internal quality control.
- Seal the plates with 8-cap strips to prevent material loss.
- Have samples sent to be sequenced at room temperature in our laboratories.
- Plate Sequencing Service: provides sequences of DNA samples resuspended in water using standard service primers. Samples in plate.
The template must consist of DNA in one of the following concentrations:
- Plasmid-DNA up to 15 kb: 100 ng / μl, final volume of 20 μl.
- Unpurified products: 100-200 bp: 5 ng / μl; 300-400 bp: 10 ng / μl; 500-600 bp: 16 ng / μl;> 1000bp 20ng / μl final volume of 20 μl.
- Purified PCR products: 3 ng per 100 bp in a final volume of 20. The DNA sample must be quantized to the NanoDrop.
- ORDER MUST BE ACCOMPANIED BY A MOD SS_01 FORM (PROVIDED BY THE FACILITY STAFF) DEBTLY COMPLETED OR PRINTED BY THE REQUEST FOR EXPLANATORY GOALS.
- Please arrange the samples in the plate in the same way indicated in the MOD SS_02 form (provided by the facility staff).
- In case the samples are > 30kbp and / or contain sections that are difficult to read, please report it in the MOD SS_01 form (provided by the facility staff). Special PCR conditions will be adopted in order to obtain the best possible sequencing results.
- The results will be sent by e-mail within the second day following the receipt of the order.
- A free repetition is foreseen in case of technical failure.
- DNA mold and sorted primers are stored for two weeks.
- Quantize the template using agarose gel or QIAXEL using a suitable MOLECULAR WEIGHT AND CONCENTRATION WEIGHT marker. The image produced by QIAxcel, on the other hand, must contain a maximum of thirteen samples, so as to allow clear interpretation and must report the default date entered by the instrumentation.
- The concentration must be normalized through the plate.
- The plates can contain different types of DNA samples arranged in columns from the first to the twelfth and from the top to the bottom.
- The size of PCR products should not vary by more than a factor of 3.
- The H12 position of the plate must be kept free for internal quality control.
- PCR products must be sent liquids in a total volume of 28 μl.
- Seal the plates with 8-cap strips to prevent material loss.
- Have samples sent to be sequenced at room temperature in our laboratories.
-Quantize the template by agarose gel or QIAXEL or by spectrophotometric reading to ensure accurate results.
• WALKING PRIMER SERVICE, SINGLE OR DOUBLE STRAND SEQUENCE WITH WALKING PRIMER.
STANDARD PRIMERS
It is possible to use the most common primers for the most frequent vectors. The complete list is shown in the table below.
CUSTOM PRIMERS
It is possible to use to perform the sequence reaction of the proper primers, following the following indications:
- the concentration of the primer must be 2 μM;
- the primer quantity must be 30 μl for the first reaction and 4 μl for each further reaction;
- the length of the primer must be between 18 and 25 nucleotides;
- the "GC" content of the primer must be equal to or greater than 50%;
- the melting temperature of the primer must be between 50 ° C and 60 ° C;
- conditions that differ greatly from what is indicated must be reported in the MOD SS_01 form (provided by the facility staff).
OLIGONUCLEOTIDES SYNTHESIS SERVICE
Oligonucleotides are short, single-stranded DNA or RNA molecules. They have a wide range of applications in biochemistry, biology, molecular diagnostics, genomics, and other molecular biology experiments. Oligos may be unmodified or modified with a variety of chemical moieties depending on their intended use, for example, the addition of 5' or 3' phosphate groups to enable ligation or block extension, respectively, labeling with fluorophores and/or quenchers for use as probes, the incorporation of thiol, amino, or other reactive moieties to enable the covalent coupling of functional molecules such as enzymes, and extension with other linkers and spacers of diverse functionality.
The CEINGE DNA LAB technology platform OLIGONUCLEOTIDES SYNTHESIS SERVICE has been working in the design and production of oligonucleotides for more than twenty years, always ensuring its users high quality and purity products.
All the molecules produced are in fact controlled without additional costs by means of MALDI-TOF, whether they are of medium size or longer or that have intramolecular and external modifications.
Oligonucleotides are supplied raw or desalted by precipitation in ethanol. At the user's request and for specific technical requirements, the oligo is supplied after purification by HPLC, reverse phase cartridges (RP-HPLC) or PAGE (Purification by Polyacrylamide Gel Electrophoresis).
The CEINGE oligonucleotide synthesis platform is able to provide the following oligonucleotide synthesis service:
DNA Synthesis Raw and desalted grade 40 nmol scale-200 nmol scale-1 μm scale
- Unmodified DNA Synthesis
- Modified DNA Synthesis
- DNA qPCR Probes
- Others According to Your Requirements
RNA Synthesis Raw and desalted grade 40 nmol scale-200 nmol scale-1 μm scale
- Unmodified RNA Synthesis
- Modified RNA Synthesis
- Long RNA Oligonucleotide Synthesis
- Labeled RNA Probes
- Others According to Your Requirements
Large Scale Oligonucleotide Synthesis Raw and desalted grade 10 μm scale
Purification by HPLC (maximum length of oligonucleotides 40 bases)
Purification by PAGE