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    QC收費標準

    昆泰銳QC收費標準

    對于您近期對于我們收費情況的疑惑,我們給您做一個詳細的解疑:

    1. 常規樣品,即樣品質量無問題,引物質量無問題的樣品,有效讀長達到800bp,即將收費。

    2. 樣品存在特殊結構的,如發卡結構,poly結構,重復序列等結構,導致反應信號衰減較快,反應讀長較短的或者結構之后出現mix套峰的,將收費。
    ???????發卡結構,存在較大和復雜的空間位阻,所以測序反應進行到此處時,容易出現信號中斷;Poly結構和重復序列,本身會形成二級結構,且Poly結構和重復序列在測序進行時,容易出現讀取堿基時出現堿基位移,所以該結構之后會出現套峰。

    3. 樣品本身質量有問題,如size較大,濃度太低,導致反應信號低,讀長較短的,讀長達到了500bp,將收費。

    4. 引物本身有問題的,如引物二級結構較多,導致測序結果反應信號很低,將收費。
    ???????較多的引物二級結構,從測序結果上反映就是在原始反應信號最前面會出現較高的引物二聚體峰,較多的引物二級結構會導致樣品在開始測序反應前,就會被引物二聚體消耗掉很多的酶和試劑,這樣可能會導致樣品在后續反應總由于酶的消耗,不足以維持后續反應,而出現反應信號很低,讀長較短。

    5. 樣品質量沒有問題,反應信號值理想,出現mix套峰的,將收費。
    ???????DNA模板上出現二處以上的引物結合位點(多引物結合位點),或者DNA模板上有嚴重的重復序列,以及測序引物不純時, 測序結果便會出現套峰現象;另外,PCR樣品或者克隆樣品出現堿基點突變或者缺失和插入突變等情況,測序結果均會出現mix套峰。

    6. 對于出現測序結果OS(反應信號值太高),LR(膠分辨率太低),DI(毛細管電泳出現延遲進樣)的情況,我們將不收費,并重新調整實驗再測序,調整后測序成功的再收費。

    7.對測序結果存在質疑的反應,需明確指出質疑的位置,若質疑區在可信區(100-700bp)之內,且無法判斷的情況,我們將提供一次免費調整實驗,若調整實驗與第一次結果一致,將收取一次測序費用。

    DNA Sequencing

    Sample Preparation Instructions

    Pricing

    • We guarantee competitive prices for both Non-profit and private sector organizations.
    • Please call in for a quote.

    Benefits

    • The most flexible sequencing service with the highest success rate
    • Skilled interpretation of your sequencing results
    • Trouble-shooting based on our expertise and our proposals to deal with sequencing difficulties (hairpin, GC-rich templates)
    • Additional services (template amplification, PCR purification) are included in the service package
    • Optional sequence editings and sequence alignment are available free of charge

    Quality

    • Average reading lengths: 700-1000 bases.
    • Color printouts of chromatographs are provided free of charge upon request.
    • Optional sequence alignment saves time when analyzing sequencing results.
    • Sequence editings are provided upon request. Sequence editing means a manual proofread performed by an experienced staff to correct mistakes /uncertainities in sequences generated by the automatic base callor.
    • Trouble shooting: We propose further trouble shooting steps based on the sequencing results. However, it is your decision whether we continue or stop the sequencing job.
    • Repeat policy: Failed samples will be repeated based on our interpretation of your sequencing results, and the repeat is free of charge.

    Turnaround

    The general turnaround time is 24 hours. Add another 24 hours for E.coli strains.

    Colony PCR

    Pricing

    • We guarantee competitive prices for both Non-profit and private sector organizations.
    • Please call in for a quote.

    This protocol is designed to quickly screen for plasmid inserts directly from E. coli colonies. The plasmid should be high copy number such as pUC18 pUC 19, or pBluescript, etc. Even though blue/white screening can be used to determine if inserts are present, this technique can be used to determine insert size and/or orientation in the vector. We have an optimized colony PCR protocol to tailor your insert size and plasmid type. After colony PCR is done, a small amount of product will be checked by by agarose gel electrophoresis. If predicted band is observed, ExoSAP process will be carried out to remove excess primer and dNTP, and then DNA sequencing will be executed.

    Template Amplification (RCA)

    Pricing

    • We guarantee competitive prices for both Non-profit and private sector organizations.
    • Please call in for a quote.

    Template amplification uses bacteriophage Phi29 DNA polymerase to in vitro amplify plasmid DNA, also called rolling circle amplification (RCA). RCA has a strong preference of circular DNA templates like plasmid DNA. Plasmid amplified by RCA is suitable for DNA sequencing. You can submit DNA samples in case you have limited DNA samples, or E.coli strains. Turnaround time for RCA amplification is 24 hours. When combined with DNA sequencing, turnaround time for the whole process is 48 hours.

    Nucleic Acid Extraction

    Pricing

    • We guarantee competitive prices for both Non-profit and private sector organizations.
    • Please call in for a quote.

    Plasmid preparation

    A plasmid preparation is a method used to extract and purify plasmid DNA. We used Qiagen Miniprep to Maxiprep kits to purify plasmid DNA for your need. We accept various forms of samples, such as bacteria culture, bacteria colony plate, bacteria seed or ligation product. After plasmid preparation is done, DNA sequencing will be preformed. Plasmid DNA or bacteria seed can be returned to you upon request.

    Genomic DNA preparation from tissue

    A customized protocol is available for extracting genomic DNA from animal tissue (commonly mouse tail). This yields DNA suitable most downstream applications, such as genotyping, Southern blotting, sequencing, etc. The genomic DNA can be returned to you upon request.

    Genotyping

    Pricing

    • We guarantee competitive prices for both Non-profit and private sector organizations.
    • Please call in for a quote.

    Genotyping refers to the process of determining the genotype of an individual by the use of biological assays. Our methods of doing this include PCR, DNA sequencing and qPCR. The technology is important in clinical research for the investigation of disease-associated genes.

    Genotyping applies to a broad range of research interests including SNP detection, transgenic/knockout determination and strain determination.

    Cosmid and BAC Sequencing

    Reactions

    Sequencing of cosmid and Bac-DNA is similar to sequencing of plasmid templates except that more DNA is needed. 2 ug Cosmid/Bac DNA is needed for direct sequencing. There is no additional charge for Cosmid/Bac sequencing.

    We also offer the whole cosmid- and Bac-DNA shotgun sequencing. Please inquire about details.

    High Throughput Sequencing Services

    Pricing

    • We guarantee competitive prices for both Non-profit and private sector organizations.
    • Please call in for a quote.

    Reactions

    Single-pass sequencing reactions are performed in 96-well PCR plates.

    Sample Submission

    • The DNA samples and the primers should be pre-mixed in defined volume.
    • Samples must be submitted in 96-well PCR plate format to preserve samples' internal order.
    • Common primers can be added by our staff free of charge upon request.

    Plasmids

    We also accept E.coli strains instead of purified plasmid DNA. We in vitro amplify plasmids from bacteria for DNA sequencing.

    PCR-products

    We purify PCR products from primers and dNTP using Exonuclease and gel filtration.

    PCR Purification

    Pricing

    • We guarantee competitive prices for both Non-profit and private sector organizations.
    • Please call in for a quote.

    We purify PCR products from primer and dNTP by using exonuclease and gel filtration columns.

    PCR products should have a single distinct band on agarose gel, otherwise, gel purification should be pursued.

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