1 Introduction To The Lab

‫بسم ال الرحمن الرحيم‬ ‫ب اْلَعاَلِمي َ‬ ‫ن‬ ‫ل َر ّ‬ ‫ي َومََماِتي ِّ‬ ‫حَيا َ‬ ‫سِكي َوَم ْ‬ ‫لِتي َوُن ُ‬ ‫ص َ‬ ‫ن َ‬ ‫ل ِإ ّ‬ ‫ُق ْ‬ ‫سِلِمي َ‬ ‫ن‬ ‫ل اْلمُ ْ‬ ‫ت َوَأَنا َأّو ُ‬ ‫ك ُأِمْر ُ‬ ‫َٰ‬ ‫ك َلُهۖ َوِب ‌َٰذِل‬ ‫شِري َ‬ ‫ل َ‬ ‫َ‬ ‫صدق ال العظيم‬ ‫سورة النعام ‪١٦٣ - ١٦٢‬‬

-This series of presentations will provide a 'step by step' guide for Molecular Microbiology Laboratory Techniques. -This course targets mainly those who are enthusiastic to contribute to science development. -So, if you have a limited knowledge in that field, hopefully by following these presentations you will be able to do your 'first' molecular experiments.

Introduction To Molecular Microbiology Lab

Usually better !

*Essentials - Lab coat. - Disposable gloves [suitable size 'S, M, L', latex or nitrile]. - Timer. - Calculator. - Set of different 'size range' pipettes (0.5-10ul, 10-200ul, 100-1000ul), plastic Pasteur pipettes. - Permanent markers, wide and fine tips. - Stationary tools [pens, pencils, eraser, scissors, glue stick, highlighters 'different colours'] for clear presentation of data in the lab book. - According to your experiment, you will need specific lab Equipments which will be mentioned later on.

-Take a tour with the person in charge or a senior lab member: to Identify your new lab. Keep small notebook : for miscellaneous information such as: * Location of cold rooms, fridges, freezers, shelves, containers (labels and contents of each). * Location of different reagents in the lab such as acids, highly flammable substances - amount allowed to keep on your bench. * Location of disposal containers for different stuff such as needles, broken glass. * Location of the glass-ware washing facility, autoclave – number and time of different cycles/day. * Names of support stuff for good communication. * Different hoods – their uses. * Other common use facilities and machines such as centrifuges – their differences. * Emergency equipments such as eye wash.

Very Important

For : - Registering every day experimental steps in detail 'ASAP'. - Loose leaf, A4 papers. -Ring binder: is the most suitable as it allows you to add any notes or lecture handout in the middle. -Each and every small notice must be included: may appear of no value at that time, but soon you will discover its importance in interpretation of experiment. -Take along the relevant textbooks which contain details about your experiment.

- Software such as 'Data base program’ can be used to keep the results.

- However, using such a programme Does NOT decrease the importance of having a lab book which must be with you in the lab while doing your experiments [outside the working space of the bench] till you are able to access a computer to put your data on. - Such a program can allow you to make a rapid search for certain information, and also considered a backup for your techniques for future use.

*Induction Course - Attendance is very important !! - Gives you information about:

- Safety measures: - 'COSHH' regulations 'Control of Substances Hazardous to Health‘ - Critical personal safety such as ‘eye safety’ when using a UV illuminator.

- Regulations of the department: - Work hours – different authorities for any queries e.g, safety. - Permission for Late hours work. - Restricted work areas, advance permission. - Lab Machines which require special training prior to individual use (e.g: Ultra-centrifuges).

Good proposal

1) Reading Literature As Much As You Can. - Takes time, but is essential before selecting a topic.

Design of your experiment - other fine tunings.

2) Accurate 'Time plan'

Regularly check if you

get an “acceptable part” of the expected results – proportional to time

3) Be ready to shift

your experiment to a

modified way in case of facing difficulty before loosing more time, but Avoid Major Change “ = New Plan”.

4) Think of availability of 'all' requirements of a good research. - Who will provide you if not commercially available. - It is better to catch them in your hand before starting the project [e.g.; control samples, antibodies for immunological assays 'Western blots, ELIZA' of new experimental product such as recombinant proteins].

of kits you intend to buy for future steps.

Read well = Good Decision

- Next day work to avoid loosing time for just preparing solutions, sterilization of instruments, or ordering stuff (already finished).

- Advance booking of equipments [such as hoods, ultra-centrifuges, freeze-dryer] which are frequently used by most of the lab members. - Register your booking details in the log book of each equipment.

- Moving

to a next step in your experiment: Do Not Do Except after

being sure of the validity of the “previous” step as long as you have the tools to check.

It’s ‘Critical’ !!!

details

Much easier - Use a wide-tip marker - Write details of the solution on the bottle or the flask

- Molecular work samples are usually contained in: Eppendorf tube - Cryovial - bijoux - Universal tube - Falcon tube.

The Most

difficult

(small surface area available for writing).

*Follow a consistent and clear way for labelling all samples: - Use fine-tip marker - write a sample number on the eppendorf cap: (Easily select the tubes you need by numbers while in the box). - On the side of the eppendorf, write: date, and a clear hint about its contents. - Specify a notebook for registering your samples' data, including the following: - Sample Number [written on the cap, and side]. - Date. - Container [for easy identification as eppendorf, bijoux, universal,...]. - Where to find it 'location' [e.g: fridge, -20 freezer, or -80 freezer, liquid nitrogen, or cold room (4°C)]. - Contents in detail.

-Send

solutions

requiring

sterilization

to

the

autoclave. [Do NOT forget to stick 'steam

indicator tape'

on the bottles or flasks to ensures efficient sterilization], OR - Use filtration according to the solution contents (e.g: if it contains sugars which are affected by high temperature). - Write your name, lab number, solution name, date, storage temperature, and shelf life on your containers before sending to the autoclave.

- Must be ready to put any order As Soon As You Can. - Keep details of each product for future use: *Product name. *Product catalogue number *Company name *Price *Delivery fees

Thank You for Attention