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    di Bernardo Lab - Systems and Synthetic Biology Lab
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    WET LAB

    Our rules

    • A- si entra sempre e solo con camice e guanti (ognuno e' responsabile del proprio camice, quindi procurateveli, scriveteci il vostro nome e assicuratevi che siano puliti, niente scuse!)
    • B- per ogni laboratorio dovete avere un camice diverso: stanza cellule e laboratorio di biologia molecolare non possono condividere un camice!
    • C- quando fate un esperimento siete responsabili del vostro bancone (o cappa) per la durata dell'esperimento e per la pulizia successiva, considerate la pulizia come parte INTEGRANTE dell'esperimento
    • D- mettete sempre il vostro nome e la data sui campioni che state preparando.
    • E - Compilate il registro cancerogeni!!! (ECCO UN LINK AL  #Resgistro# )

     



    Plasmids & Cells

    Instructions for plasmids' map and sequence

    • All the plasmids should be kept in GENBANK format. This format can be read by any software and allows to easily annotate all the sequences.
    • All the plasmids you use, can be downloaded in GENBANK from ADDGENE.
    • Once in ADDGENE you found your plasmid, click on 'Sequence' under 'Plasmid Links', then on 'Analyze Sequence' and from here you just click on 'GENBANK'.
    • Copy the sequence and paste it in TextWrangler (MAC) or Notepad (PC) and save it a txt plain text (not WORD or RTF format) in TextWrangler click on CONVERT TO ASCII
    • Now you can view and modify your plasmid with SerialCloner/GENIOUS or any other program

    Perform in-silico cloning using the following software

    Here you can find a tutorial on how to carry out in-silico cloning experiments

    Enzymes Inventory

    Here you can find the restriction enzymes

    The Plasmid and Cells Database

    All of the cells and plasmids must be stored in our database (Click here for instructions).



    FACS Analysis tool for Matlab

    PDF TUTORIAL

    In \data\wetlab\FacsAnalysis you will find a tool to analyze facs data with matlab.

    Instructions:

    • In matlab environment move to ...\data\wetlab\FacsAnalysis\ folder by typing "cd" followed by the entire path where the tool is in
    • Type in the command line: "FacsAnalysis" to start
    • Now you can import a facs file (.fcs 3.0 format), define a polygon to select the cells, plot and save data (.mat and .xls extensions are allowed)
    • A complete tutorial is available here, for any further information please contact Gianfranco

     

    Supply centre by Life Technologies - How to order

    PDF Tutorial

    Oligonucleotides' synthesis by Sigma - How to order

    • Fill in the excel file orders form indicating your name as "Reference", the project number and Sigma as Providing Company"
    • email a.calicchio@tigem.it with the form attached asking for the confirmation number
    • enter the confirmation number in the form (before the project number)
    • order online on the Sigma website and wait for the order confirmation
    • email gagliardi@tigem.it with the order form and the order confirmation (from Sigma) attached
    • save the two files in the "Orders" folder in Data for backup

    Nikon Eclipse - How to use it

     

    Filters mounted on the microscope

    Name: YELLOW GFP BP HYQ

    Ex 490 - 510
    Dm 515
    Em 520 - 550

    cod. MXU96211

    Pos. 4

    Use: YFP, Venus

    Name: UV-2A (DAPI)

    Ex 325 - 375
    Dm 400
    Em 420

    Pos. 5

    Use: Dapi

    Name: CYAN
    Ex 426 - 446
    Dm 455
    Em 460 - 500

    cod. MXU96210

    Pos.UN-MOUNTED

    Use: cyan

    Name: PSTN GFP

    Ex

    Dm

    Em

    cod. c128752

    pos: 2

    Use: green

    Name: TRITC HYQ

    Ex 530 - 560
    Dm 570
    Em 590  - 650
    pos.3
    Use: red dye and cherry

    Name: FITC

    Ex 465-495

    Dm 505

    Em 515-555

    Pos.1

    Use: GFP, Venus, YFP


      Plasma Cleaner

      General Infos

      PDMS bonding process

      Barbara, Lorena and Giansimone have been trained to use it, for enquiry contact Gianfranco


      Inventory


      Confocale

      Linee guida


      Reference Material

      • Microfluidics
      1. Fabrication of a microfluidic device, an example (Video)
      2. Microfluidics (wikibook)
      3. PDMS a review



        Protocols