Sequencing Order Form (Kindly send the order form along with the samples)

Transcription

1 Sequencing Order Form (Kindly send the order form along with the samples) Bioserve Biotechnologies (I) Pvt. Ltd. Date: From: Billing Address: Distributer Details: Name: Address: Bill To: Address: Name: Address: -id: Phone: -id: Phone: -id: Phone: Note: This form will be treated as a work order. Filling up of all the details is mandatory. Send hard copy of page no. 1, 5 & 6. (Kindly read all the instructions before filling up the form and do not edit the order form ) Signature For Office Use Only: BBIPL/SEQ/ - / Date Received: P a g e 1

2 Mandatory Fields: Bioserve follows the protocol optimized by ABI for sample preparation. Template DNA supplied should satisfy the following criteria : Salt free RNA free and EDTA free Template DNA must be finally re-suspended in deionized water only. Quantity (and concentration) of template DNA required for sequencing service: Table 1: Prepared Template and Primer Submissions: Template Type Template Size Template (ng/µl)* Primer(pmols/µL)* Plasmid <20kb PCR product <1000bp PCR product >1000bp Large Constructs (BAC) >20kb * Additional Volume of 5µl for each Additional reaction. * Sample concentration in 1µl should be nearer to the internal standard shown in the above table, when visualized on 1% agarose gel. * Quantity of samples for gel elution should be 100µl. P a g e 2

3 Criteria for primers used in sequencing: Primer sequence selection, synthesis and purification have significant effect on the quality of sequencing data. We recommend the following criteria : 1. High Purity (either HPLC or Desalted) Primers. 2. Primers bases long usually give good specificity. Long Primer (>35mer) is not preferred 3. GC content of 50% to 55% is preferred. 4. No mismatch is present and no alternative hybridization sites are present in the template. 5. RAPD primers (10 mer) cannot be used for cycle sequencing reaction due to low annealing temperature. 6. For cycle sequencing, primer with melting temperature between 50 C to 60 C is recommended. Avoid low melting temperature (40 to 45 C.) Melting temperature (Tm) is calculated as 4(G+C) + 2 (A+T). No secondary structure particularly at the 3' end should be present. Primer Walking Service: 1. This service includes DNA extraction, purification, Primer designing, synthesis, desalting, sequencing and assembling of the final sequence. 2. Information & sequence report will be provided to the customer at each stage of the sequencing by primer walking. 3. In case of unforeseen problems due to GC/AT rich regions of template DNA, the customer will be informed for suggestions and billed according How To Send Samples: to the number of reactions performed/primers synthesized. 1. DNA must be transported in dry ice / coolant packs, at sufficiently low temperature to prevent degradation. DNA in solution should not be sent at room temperature. 2. Customers are requested to send the printed Order Form (page no. 1, 5 &6) filled in along with the and contact number. 3. Please submit your template and primer in 1.5ml Eppendorf tubes and be sure to label your tubes as in the order form. Tubes submitted in other formats will not be accepted. 4. Samples received in lab after 2PM will be considered as the next day order. 5. We routinely store samples for 20 days unless instructed otherwise. P a g e 3

4 Result of DNA Sequencing Service: 1. Bioserve will provide data by -of each sample in the form of Electropherogram/chromatogram along with the internal sequencing control. 2. The sequencing read length provided will be up to 600bases. 3. Result depends on the successful fulfilling of quality and quantity criteria. 4. The Sample will be charged for <600bps also, if our positive control gives a good intensified read length. We do not re-run the failed reactions unless requested explicitly by the customer through within 3days of receiving the data. The extra run is charged. Delivery time: 1. Sequencing service: Approximately 3days. A] PCR products and Plasmid DNA- 3working days B] PCR products with purification-4 working days C] Plasmid DNA Isolation and sequencing-4 working days 2. Primer walking services: Approximately 1 week for every 1 kb. P a g e 4

5 Details of DNA templates for sequencing S. No Sample ID Type of sample (Plasmid/PCR product/ssdna/ stab cultures) Medium (Deionized H2O/Lyophilize d) Volume of Medium Min (10-50 l) If Lyophilized, Qty of sample Conc. Of DNA Template ng/ l Construc t Size Insert Size Vector Type Is it GC/AT Rich/Lon g repeats/ Purificati on protocol used P a g e 5

6 Primer Detail Form S.No (Corresponding to template) Primer to be used (Universal M13/T7/T3/SP6/ Custom) Tm of Primer Annealing Temp. used by scientist Conc. Of Primer (pm/ l) Volume of medium Qty of Primer Is Primer Purified Length of Primer with sequence P a g e 6

7 Primer Walking Services Initial primers to be sequenced with : Sequencing to be done for : One strand / both strands Bank Details of BIOSERVE BIOTECHNOLOGIES (INDIA) PVT LTD, HYDERABAD 0.1 Name of the Bank State Bank of Hyderabad (SBH) 0.2 Branch Name/Branch Code Habsiguda Branch/ Branch Address Uppal Road, Hyderabad Ranga Reddy District, A.P Digit MICR No. of the bank/branch as appeared on the cheque Account Type (Savings/Current/Cash Credit) Cash Key Account 0.6 Account Number (as appeared on Cheque Book) IFSC Code of the Branch S B H Y Information for Cheque Deposit P a g e 7