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DNA Sequenceing Introduction
 

 DNA samples should be delivered to Rm401-5 before 9:30 every morning with a complete sample sheet. The DNA sequencing service will be provided at DNA Sequencing Core Facility of IBMS.

 
DNA Sequenceing Equipment Contact: Dia-Nan Lin   Tel-9527      
 

 

Equipment

Models

Applications

Link

 

ABI 3730XL DNA Analyzer

( located in the DNA Sequencing Core at IBMS )


ABI PRISM

DNA Sequencing

DNA Sequencing  WORK REQUEST

Sample preparation

chromassoftware download

 

 

 
DNA Sequenceing Regulations Contact: Dia-Nan Lin   Tel-9527    
 
Submit sample

1.

The quality of plasmid samples should be determined by 260/280 nm ratio. Generally the ratio should be approximately 1.8 for pure DNA.

2.

DNA samples should be delivered to Rm401-5 before 9:30 AM

3.

 Total volume of your sample should be 12 λ in a 0.2 ml PCR tube
※If using our primer ( M13 Forward, M13 Reverse,T7 Promoter, SP6 Promoter, T3 Promoter), the total volume of your sample will be 10 ul in a 0.2 ml PCR tube. Please indicate the primer on the back of the form. 


DNA type

Concentration required
(Use Core Facility primer)

Concentration required
(Use your primer)

Total volume

10 μl
(DNA only)

*12 μl
(DNA+Primer)

Plasmid/dsDNA

0.5 μg

0.5 μg

Single-stranded DNA

62.5-125 ng

62.5-125 ng

PCR product

Size 200-500 bp: 7.5~ 25 ng
Size 500-1,000 bp: 12.5~ 50 ng
Size 1,000- 2,000 bp: 25~ 100 ng

Size 200-500 bp: 7.5~ 25 ng
Size 500-1,000 bp: 12.5~ 50 ng
Size 1,000- 2,000 bp: 25 ~ 100 ng

Your custom primer

-

4~ 10 pmole (40 ng),
volume:2 μl .

 

4.

Core lab primer ( M13 Forward, M13 Reverse, T7 Promoter, SP6 Promoter, T3 Promoter )


Primer supported by Core Facility Laboratory

Primer type

Primer sequence

21M13 Forward Primer

5’  GTT TTC CCA GTC ACG AC  3’  (17 mer)

21M13 Reverse Primer

5’  CAG GAA ACA GCT ATG AC  3’  (17 mer)

T7 Primer

5’ TAA TAC GAC TCA CTA TAG GG  3’ (20 mer)

SP6 Primer

5’ TAT TTA GGT GAC ACT ATA G  3’  (19 mer)

T3 Primer

5’ CCA TTA ACC CTC ACT AAA GG  3’ (20 mer)

 

5. Strip tubes: If the number of samples is 8 or more, please submit your samples in 8-strip tubes.96-well plate: If the number of samples is over 48, you may submit your samples in a 96-well plate that is well sealed with aluminum foil tape.
  For additional information, please refer to Sample preparation
   
 
DNA Sequenceing Procedures Contact: Dia-Nan Lin   Tel-9527    
 
Service object
 

We provide DNA sequencing services to all ICOB researchers and their collaborators.

   
Usage protocol
  Users: Submit--Prepare samples and fill out aDNA SEQUENCING WORK REQUEST  
   ↓   
  Deliver to Rm401-5  
   ↓   
 

Core: Collect samples for DNA sequencing on the ABI 3730XL DNA Analyzer at IBMS

 
   ↓   
  Data will be delivered by e-mail on the next work day.  

 

 

 

 

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