Sequencing Service Order

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Though the all the sequencing procedures can be done without following the guideline, we highly recommend sending the samples according to the sample requirements.
Sequencing Service will be processed after customer samples are received, so separate settlement must be made separately from other products for the convenience of settlement
Order manual / How to check data

Sequencing Service Needed

Quantity

Custom Sequencing Primer Synthesis [help]

Quantity

Additional Service Needed [help]

Quantity

Sample Type

Total No. of Reactions

Storage Type

Contact Person Name

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Reaction Information [help] [Search my orders]

		Sample			Primer
	Tube	Name	Vector name	Size(bp)	Name	Conc.(pmol/ul)

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Download Form(Plate) Download Form(Tube)

Sequencing Service Order

In stock

$0.06

Summary

Product Overview

Overview

Our company not only utilizes advanced automatic gene analyzer ABI3730XL capable of sequencing various templates such as plasmid DNA and PCR products, but also consists of experts and rich infrastructures to provide reliable data for researchers. Furthermore, using our own gene synthesis techniques, both sequencing and oligo synthesis service can be simultaneously run to get the results in a short time. This is especially the case for primer walking and virus/vector sequence validation.

Features and Benefits

• Accuracy

High quality data with accurate results through automated process (Phred Score ≥20, Guaranteed read lengths ≥700bp)

• Free universal primer

Supply of commonly used universal primer capable of being selected upon order

Workflow

• Working Day: Completion within 48 hours of receiving time

• After completing the service, we will provide 4 types of files (ab1, text, PDF, full text) and reports.

• Samples and primers will be discarded after 1 month. A seperate request is required for a long-term storage

▶ Consultation

E-mail sequencing@bioneer.co.kr

Literature/Support

◈ Brochure

• All about Bioneer's Custom Services

◈ Quality Assurance

Bioneer is the holder of Quality Management System Certificates for the following standards.

• ISO 9001 – certificate

Ordering Info

Sequencing service list

▶ Sequencing service list

• Standard Sequencing

Standard Sequencing service analyzes the desired Plasmid/PCR product region using a primer. It is optimized for most of the normal samples.

• Full Length Sequencing (Primer walking)

Full Length Sequencing service analyzes plasmids, long inserts, or PCR products by primer walking. It takes about 1 week in 3 kb. An additional period will be required depending on the size.

After the service is completed, the data is provided in the form of contig file after each single data and assembly. The Primer information is written in the report and uploaded with the results.

• Difficult Sequencing

Difficult Sequencing service aims to analyze templates difficult to be analyzed with standard sequencing such as those having GC/AT rich regions, repeated sequences, secondary structures, etc.
This service is also optimized for Bioneer's long time experiences and technique that utilizes analysis conditions and reagents different from standard sequencing.
Additional service list

▶ Additional service list

Depending on the requested samples, additional services can be chosen.

* All material is serviced by our products.

1) Genomic DNA → PCR → Non-purified PCR product → PCR purification/Agarose gel extraction → Purified PCR product

2) Genomic DNA → Primer design → PCR optimization → PCR → Non-purified PCR product → PCR purification/ Agarose gel extraction → Purified PCR product: Custom Sequencing

• Purification

A purification process must be done to remove impurities prior to the direct sequencing step of PCR products. This is especially crucial when high-quality data is required to eliminate non-specific bands.

- PCR purification:

This method uses a spin-column method, which is suitable when it is certain that no non-specific band will occur. This service uses our PCR Purification Kit PCR Purification Kit (AccuPrep^® PCR Purification Kit).

- Agarose gel extraction:

This method extracts DNA fragments from an agarose gel to acquire only the desired bands, which is suitable when non-specific bands occur in the PCR product. This service uses our PCR Purificaiton Kit (AccuPrep^® Gel Purification Kit).

• PCR & Purification

If you send the genomic DNA and specific primers, along with background information such as optimized PCR conditions, product size, etc., we can confirm the size to undergo the suitable purification process after PCR. Primer synthesis service is also available if needed. This service uses our high-fidelity PCR premix (AccuPower^® ProFi Taq PCR PreMix)and PCR Purification Kit.

Sample preparation condition

▶ Sample preparation condition

• Sample Preparation

Type	Concentration	Volume/rxn	Purity
Plasmid DNA	≥100 ng/µl	≥10 µl	A₂₆₀/A₂₈₀: ~1.8 A₂₆₀/A₂₃₀: 2.0-2.2
PCR product	≥50 ng/µl	≥10 µl
Non-purified PCR product	≥ 50 ng/µl	≥20 µ
Genomic DNA	≥ 30 ng/µl	≥30 µl

• Primer Preparation

Type	Concentration	Volume/rxn	detail
Universal primer	Free primers are available to use
Specific primer	5 pmol/µl (5 µm)	5 µl	Sequencing primer titration conditions: 18~22 mer, TM 50~56°C
Ordered primer	Sequencing and oligos can be ordered at the same time.

• Sample Enclosure

- Single tube type

When preparing with a 1.5 ml microcentrifuge tube, please specify the sample name as it appears on your order sheet.

- Plate type

When preparing with plates, it is highly recommended to use 96-well plate with V-type bottom with samples placed in the order as shown in the diagram below and seal tightly using a cap to prevent cross-contamination.

Service price

▶ Service Price

Cat. No.	Service Description	Unit	Price ($)	Note
Sequencing Service
S-3010-1	Standard sequencing	<96 rxns	6.00/rxn
S-3010-2	Standard sequencing	≥96 rxns	5.00/rxn
S-3010-6	Full length sequencing	kb	6.00/kb
Additional service
S-3010-3	PCR purification	ea	2.00/ea
S-3010-4	Gel-extraction	ea	3.00/ea
S-3010-7	Custom Sequencing Primer Synthesis	mer	0.29/mer	Based on 25 nmol

Free Primer

▶ Free Primer

Primer name	Sequence (5' → 3')	Length [bases]
T7promoter	TAA TAC GAC TCA CTA TAG GG	20
T7terminator	GCT AGT TAT TGC TCA GCG G	19
T3	AAT TAA CCC TCA CTA AAG GG	20
SP6	ATT TAG GTG ACA CTA TAG	18
EBV-R	GTG GTT TGT CCA AAC TCA TC	20
BGH-rev	CTA GAA GGC ACA GTC GAG GC	20
M13F(-40)	GTT TTC CCA GTC ACG AC	17
M13R(-40)	CAG GAA ACA GCT ATG AC	17
M13F(-20)	GTA AAA CGA CGG CCA GT	17
M13R(-20)	GCG GAT AAC AAT TTC ACA CAG G	22
pGEX5	GGC AAG CCA CGT TTG GTG	18
pGEX3	GAG CTG CAT GTG TCA GAG G	19
pQE-forward	CCC GAA AAG TGC CAC CTG	18
pQE-reverse	GTT CTG AGG TCA TTA CTG G	19
EGFP-C	CAT GGT CCT GCT GGA GTT CGT G	22
EGFP-N	CGT CGC CGT CCA GCT CGA CCA G	22
RVprimer3	CTA GCA AAA TAG GCT GTC CC	20
RVprimer4	GAC GAT AGT CAT GCC CCG CG	20
GLprimer1	TGT ATC TTA TGG TAC TGT AAC TG	23
GLprimer2	CTT TAT GTT TTT GGC GTC TTC CA	23
CMV-F	CGC AAA TGG GCG GTA GGC GTG	21
CMV30	AAT GTC GTA ATA ACC CCG CCC CGT TGA CGC	30
CMV24	TAT TAG GAC AAG GCT GGT GGG CAC	24
Gal4AD	TAC CAC TAC AAT GGA TG	17
Gal4BD-F	TCA TCG GAA GAG AGT AG	17
Gal4BD-R	TTT CTT TGG AGC ACT TGA GC	20
MATCHMAKER3	GTG AAC TTG CGG GGT TTT TCA GTA TCT ACG AT	32
pBAD-For	ATG CCA TAG CAT TTT TAT CC	20
pBAD-Rev	GAT TTA ATC TGT ATC AGG	18
SV40-pArev	CCT CTA CAA ATG TGG TAT GG	20
SV40-pAF	AAA TAA AGC AAT AGC ATC AC	20
malEF	GGT CGT CAG ACT GTC GAT GAA GCC	24
pCold-F	ACG CCA TAT CGC CGA AAG G	19
pCold-R	TCC CCG CCA AAT GGC AGG GA	20
3AOX	GCA AAT GGC ATT CTG ACA TCC	21
5AOX	GAC TGG TTC CAA TTG ACA AGC	21
pBabe-F	TGA CCT GGG AAG CCT TGG CT	20
pBabe-R	TTG CTG ACT AAT TGA GAT GCA TGC TTT	27
V5_Reverse	ACC GAG GAG AGG GTT AGG GA	20
pJET1.2F	CGA CTC ACT ATA GGG AGA GCG GC	23
pJET1.2R	AAG AAC ATC GAT TTT CCA TGG CAG	24
27F	AGA GTT TGA TCM TGG CTC AG	20
344F	CCT ACG GGA GGC AGC AG	17
518F	CCA GCA GCC GCG GTA ATA C	19
785F	GGA TTA GAT ACC CTG GTA	18
519R	GWA TTA CCG CGG CKG CTG	18
800R	TAC CAG GGT ATC TAA TCC	18
907R	CCG TCA ATT CMT TTR AGT TT	20
1492R	TAC GGY TAC CTT GTT ACG ACT T	22
ITS1	TCC GTA GGT GAA CCT GCG G	19
ITS2	GCT GCG TTC TTC ATC GAT GC	20
ITS3	GCA TCG ATG AAG AAC GCA GC	20
ITS4	TCC TCC GCT TAT TGA TAT GC	20
ITS5	GGA AGT AAA AGT CGT AAC AAG G	22
LR0R	ACC CGC TGA ACT TAA GC	17
LR5	ATC CTG AGG GAA ACT TC	17
LCO1490	GGT CAA CAA ATC ATA AAG ATA TTG G	25
HCO2198	TAA ACT TCA GGG TGA CCA AAA AAT CA	26
pET-Upstream	ATG CGT CCG GCG TAG A	16
pBA-F	ATT GTC TCA TGA GCG GAT AC	20
pBH-R	GCG TTA TCC CCT GAT TCT GT	20
pBIC_F	CTC ATG AGC GGA TAC ATA TTT G	22
pBIC_R	GCC GCA GCC GAA CGA CCG AG	20

FAQ

▶ Receipt

What do I need to prepare to use this service?

After placing your order, you must prepare the sample and the primer and send them to us.

How do I place the order online?

Please refer to the ordering information.

What kinds of universal primers are provided by Bioneer?

Please refer to the free primer list.

Can I order oilgos for sequencing?

Yes you can. You can order them together on the Sequencing order page. After their synthesis, they will be used immediately for the sequencing.

Can you provide more information on shipping methods?

Please send the samples to the address below.
(Address: 8-11, Munpyeongseo-ro, Daedeok-gu, Daejeon 34302, Republic of Korea | BIONEER Genome Analysis Team)

What is a re-reaction service?

Re-reaction service provides one more sequence check for samples and primers for those have been previously analyzed, but require another run for further improvements of sequencing results.
Data such as sequencing reports can be recorded and delivered for free. After receiving the first report, the re-analysis results can be obtained within two days.

What is the difference between additional analysis and re-reaction service?

Re-reaction, as explained above, uses the same sample and primers for another analysis. While our company guarantees at least 700 base readings, we will provide one more sequencing service for free if the readings are less than that.

On the other hand, additional analysis service provides another sequencing for the same samples, but using different primers. Thus, additional analysis will cost a service fee.

I would like to request an additional analysis.

In this case, as we need to keep checking the remaining amount of samples, you must write your initial receipt number in the comment section or select "Previous Order Sample Search" when entering the reaction information.

I would like to request sequencing using the primer I have used before.

You must have requested to store the used primers separately at the time of the order. Otherwise, they would be discard after 1 month.

To request sequencing using the stored primer, you can select the check box in the order page.

▶ Result

How many read lengths can I get through the sequencing service?

We guarantee 700 bp for single extension, but we usually get about 1.2 kb of the reading. If you request both direction analysis, you can receive high quality data up to about 1.4kb.

How is the results provided?

Reports will be provided in three formats (ab1 / seq / PDF file) with seq merge text file and reports per single extension.

- Ab1 file: AS electropherogram file viewable with viewer program

- Seq file: Base calling text file

- PDF file: Ab1 file converted to PDF format

- Seq merge text file: File combined with analyzed sequence of all the requested samples into a text file

- Report: A report that contains information and forwarding information on how to analyze and improve results

What do A, B, C, and D group imply on the sequencing report?

Results in low quality are categorized into four groups depending on the data aspects for easy understanding.

Group A) Data with low QV values which may be improved through re-analysis which can be done free for once.

Group B) No electropherogram; this case may be made from the analyst's mistake, so we will undergo re-analysis free for once.

Group C) Sequencing limited by the DNA sequences; re-analysis service will not be available as improvements cannot be made.

Group D) Other causes such as primer mis-matches; re-analysis service will not be available as improvements cannot be made.

▶ Payment

Where can I contact for payment inquiries?

We support online payments. For more information, please contact Customer Support Center (Tel. +82-42-930-8777) and we will kindly reply you.

Can I get the printed quotation?

You can simply print it out as you make your online order. Contact our Customer Support Center (Tel. +82-42-930-8777) if you want to issue the quotation separately.

In case of mass ordering, price adjustment is possible. Please consult with your salesman.

▶ Etc

How long will the analysis take?

Send the result within 48 hours when the reception is completed.

What is the retention period for the samples requested?

Samples are kept for one month after the order date.

The primers can be kept, but Bioneer is not responsible for degradation caused by long-term storage. It is recommended to freeze in several tubes and change it every 1 month for long-term usage.

How can I use the additional services?

You can choose the desired service according to your requesting sample type when ordering through our online page.

Is it possible to collect the remaining sample after analyzing the results?

Yes you may.

May I ask for the solutions and the instruments your company use for the analysis?

Refer to the information below.

▶ Sequencing Process Information

1) Material & Instruments

① AllInOneCycler™ 96 well PCR system (Block PCR Instrument, Bioneer)

② BigDye^® Terminator V3.1 sequencing kit (Applied Biosystems, USA)

③ ABI3730XL (Applied Biosystems, USA)

2) Methods

① Sequencing PCR Cyclic

<Refer to the table below for the amount of template used in PCR>

• DNA size/ DNA amount

Template	DNA amount
PCR product
100~200 bp	5~10 ng
200~500 bp	10~20 ng
500 bp~1 kb	20~50 ng
1~2 kb	50~100 ng
≥2 kb	100~200 ng
Double strand	200~500 ng
Cosmid, BAC	300~600 ng
Genomic DNA	2~3 µg

② Purification

③ ABI3730XL Running

Sequencing Service Order

Overview

Features and Benefits

Workflow

◈ Brochure

◈ Quality Assurance

▶ Sequencing service list

▶ Additional service list

• Purification

• PCR & Purification

▶ Sample preparation condition

• Sample Preparation

• Primer Preparation

• Sample Enclosure

▶ Service Price

▶ Free Primer