Thursday, September 16, 2010

Protocol Gold PCR


PROTOCOL PCR GOLD

1.      prepare product sample ekstraksi, then contents form pcr. count sample and make mastermix for sample total augmentings control negative n add 1 to eror pipetting. our for example has sample 5, add control negative 1 and eror 1 so we make mastermix to 7 tubes. if has which are positive result, we also can add control positive. add 1 eror to every multiple 10.2. prepare all reagen (h2o, buffer pcr gold (yellow), dntps, mgcl2,  and primary forward and revers
2.      take 1 tube strip complete with enclosing it. give initial label, date,  and primary. then write number 1 until 8 in tube neck strip. prepare 1 tube eppendrof 0,5 ml and give label mm (master mix)
3.      now, we liquefy all reagen (usually kept at frezer, certain it frozen, so it necessary liquefy)ed with hold tight and ascertain all melt finely. shakes to ascertain there is no component endap mendasar tube.
4.      before memipet reagen, always mix beforehand by using pipette.
5.      take h2o appropriate count that we can,  and input to tube mm (volume appropriate protocol)
6.      furthermore, input buffer pcr, dntps, mgcl2, primary 1 and 2, with enzyme taq gold (take with pipette kusus enzyme and soon return to frezer the enzyme, watch out expensive goods)
7.      then share 24 µl mm into each tube hole strip that we are label,  and input dna template result ekstraksi as much as 1 µl, don't forget at mix each time insert dna.
8.      to assure mixing and cause the loss of bubel, so try at spin.
9.      furthermore, close up tube strip that full dna and mm,  and input intoes thermocycler. running engine with program gold.
10.  wait up to end process,  and neat dehhh…. congratulation wait the result…. .

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