GENETIC ENGINEERING - Sequencing

Biomedal has at your disposal several DNA sequencing services, offering high quality reactions up to 700 bp, with a fast reply time and competitive prices for all your sequencing needs.

If you need a simple sequencing (PCR/plasmids) or one that is more complex, like for cosmids, phagos and BACs, Biomedal offers a complete and efficient service.

Moreover, we give alignments and comparison of capillary electrophoresis sequences, just as Primer-walking service, available for sequences up to 15 kb.

General conditions of the service:

Samples sending:

- DNA and primers must be sent in separate tubes.

- In solution or dehydrated, EDTA free.

- Refrigeration is not necessary. (NOTE: If you send the sample refrigerated do not use ice).

- Isolate the tubes with parafilm and wrap in padded material.

- Write FRAGILE MATERIAL on the envelope.

Specifications and conditions for DNA sequencing:

Service characteristics:

- For plasmics, PCR product, cosmids, phagos and BACs.

- Reading up to 700 bp, its depends on the quality and the DNA mold.

- Design and synthesis of specific primers.

- Alignment and comparison of sequences (optional)

- Store the DNA mold and primers during one month.

- Quality analysis of each sequence (occuracy > 99%).

- Universal primers offered.

Your sample:

To obtain the best result, send your DNA samples highly purified. Samples and primers must be provided in separate tubes.

The minimum quantity of DNA necessary for the sequencing are:

- Plasmids: 100-200 ng/µl in ddH2O, 1µg/reaction.

- PCR products: 30-60 ng/µl in ddH2O, 1µg/reaction.

- Cosmids and phagos: minimum of 400 ng/µl, 3,0 µg minimum/reaction.

- Primers: minimum of 10 pmol/µl in ddH2O in a total of 50 pmol minimum/reaction.

Minimum volume recommended for send DNA samples: 15 µl of ddH2O.

PRIMERS FORWARD	SEQUENCE
T7 FwdpGEM	5'-GTA ATA CGA CTC ACT ATA GGG C-3'
T7 Fwd	5'-TAA TAC GAC TCA CTA TAG G-3'
M13 Fwd	5'-TGT AAA ACG ACG GCC AGT-3'
pT7-ET	5'-ATG CGT CCG GCG TAG A-3'
nT7fwd	5'-AAY ACG ACT CAC TAT AG-3'
PQE30fwd	5'-CCC GAA AAG TGC CAC CTG-3'
5pGEX	5'-GGG CTG GCA AGC CAC GTT TGG TG-3'

PRIMERS REVERSE	SEQUENCE
T7 Rev	5'-GCT AGT TAT TGC TCA GCG G-3'
T3	5'-ATT AAC CCT CAC TAA AGG G-3'
M13 Rev	5'-CAG GAA ACA GCT ATG ACC-3'
SP6	5'-ATT TAG GTG ACA CTA TAG-3'
BGHrev	5'-TAG AAG GCA CAG TCG AGG-3'
PQE30rev	5'-GTT CTG AGG TCA TTA CTG G-3'
U19mer	5'-TTT TCC CAG TCA CGA CGT-3'
3pGEX	5'-CCG GGA GCT GCA TGT GTC AGA GG-3'

Specifications and conditions of capillary electrophoresis service:

If you have your DNA samples amplified and only want based readings, we offer the reading service using capillary electrophoresis.

Note: This service only goes for samples sequenced by BigDyeTerminator v1.1 and v3.1 Kit of Applied Biosystems.

Characteristics of the service:

- Reading up to 700 bases

- Reactions of electrophoresis making by the customer.

Your sample:

The material amplified and purified must be given in a dry pellet.

Specifications and conditions of Primer-Walking service:

If you have large sequences that could not be read in a simple reading, Biomedal offers the Primer-Walking service, for one-way or two-ways reading.

Service characteristics:

- Reading up to 15 Kb.

- Design and synthesis of specific primers.

- Alignment and correction of sequences.

- Store the DNA mold and primers during one month.

Ref.	Product
SE-3384	Plasmid Sequencing 1-2 services
SE-3385	Plasmid Sequencing 2-8 services
SE-3386	Plasmid Sequencing 9-49 services
SE-3387	Plasmid Sequencing = o >50 services
SE-3388	PCR Sequencing 1-2 services
SE-3389	PCR Sequencing 3-8 services
SE-3390	PCR Sequencing 9-49 services
SE-3391	PCR Sequencing = 0 >50 services
SE-6667	Cosmid/Phage/BAC Sequencing