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Post PCR

By Bertha Butler,2014-12-24 15:21
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Post PCR

    Post PCR

    General

    ; Please use only your groups consumables i.e. agarose, tips, TBE buffer,

    reagents, etc. Non shared consumables are usually marked with your

    group name.

    ; Please don’t take any equipment from pre PCR into post PCR and vice

    versa - equipment must be kept in the designated lab.

    ; Please don’t waste autoclaved tips for gel loading. There are refillable

    (unautoclaved) tips for this very purpose.

    ; Please make sure you have turned the heat sealer off if you are the last

    person needing it.

    ; Please empty the clinical waste bin when it is full - use a cable tie to seal

    the bag, write “Post PCR Lifeline” on the bag with a permanent marker,

    dispose of it in the clinical waste bin outside the building (the key for it is in

    a drawer in the cell culture room ask if unsure). Replace by double

    bagging the clinical waste (yellow/orange) bags in the bin. ; Please tidy up after yourself by throwing away used tissue, gloves,

    replacing anything that you might have used up and packing equipment

    away that you have used, so that someone else doesn’t have to clear up

    your mess before starting their work.

    ; Please make sure the scalpels are back in the polystyrene holder after

    use.

    The PCR Tetrad

    ; Please use the rota to book the block(s) you want to use and specify the

    correct time you need. Make sure when using the tetrad you use the

    actual block you have booked on the rota (see the front of the tetrad for

    the block positions).

    ; Please ensure that you have set your “forever” step to 15 ?C instead of

    4 ?C and try not to leave your samples on overnight if you can prevent it

    (this prolongs the life of the machine).

    ; Please end the PCR program you were using once your samples are

    removed from the machine by pressing proceed (every program has an

    “END” step, without ending the block will continue the cycle).

    ; Please make sure that you have turned the whole PCR machine off when

    it is not in use.

Gel Preparation

    ; Please use the designated ethidium bromide pipette in the fume hood for

    pipetting of ethidium bromide ONLY and never for anything else (for health

    and safety reasons).

    ; Please discard tips used with ethidium bromide straight into a sharps bin

    and not into any intermediate waste pots.

    ; If you use the last of the 5 X TBE then make some more!

    ; Please clean up any TBE buffer that you spill as soon as possible as the

    TBE salts are very difficult to remove from bench surfaces once dry. ; Please rinse the gel combs after every use as it prevents “little spots”

    developing on your gels.

    ; Please don’t leave the gel trays in the TBE buffer unnecessarily and rinse

    clean after every use.

    ; If you don’t use all of the gel that you have made, either leave a note with

    the percentage gel and date made on the gel tank for someone else to

    use or discard it (it is a health and safety issue leaving unused gels lying

    about on the bench).

    ; Please replace the TBE buffer in the gel tank after every 2nd time of using

    it. We will start a system where you tick off on a piece of tape on the gel

    tank every time you have used it.

    Gel Doc

; When using the gel doc PC ensure you wear gloves at ALL times.

    ; Please don’t leave your gel in the gel doc unnecessarily. Take care when

    turning on the UV lamp freeze the image once it is at the right intensity

    and turn the lamp off straight away the computer will keep the image

    once you have frozen it. Make sure you turn the UV lamp off once you are

    finished the bulbs are VERY expensive to replace.

    ; Please wipe the gel doc surface after every use and ensure it is switched

    off.

    ; Please switch off the gel doc printer before shutting down the PC,

    otherwise it re-prints the last image captured and wastes the printing film. ; If you are the last user of the gel doc system of the day please shutdown

    the PC - the computer has to be switched off manually as it is very old!

    If in doubt about using any of the equipment please do not hesitate to ask

    for help from others in the lab or if no one else is around call ext 6181 and

    ask Kay Mehers or Berendine van Zyl.

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