Polymerase Crap Reaction

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Sangat aneh bila kamu masukkan sejumlah campuran bahan kimia dalam satu tiub kecil, masukkan dalam mesin yang sepatutnya tambahkan lagi isipadu, tapi bila semuanya…teeet teeeet teeet tamat…yang kamu dapati..eh kenapa kosong ni? T_T

 

Jadi, kata prof saya, tutup rapat-rapat, penyet-penyet atas meja.

 

Saya dah buat, prof.  -_-”          Saya tahulah saya lembik.

 

Saya pun terus buka buku “WORKING WITH DNA, Metzenbergghh”

“Heated lids may soften the top of the thin-walled tube and cap, and allow leakage of the pressurized water vapor during the high-temperature denaturation steps.”

Kemudia google dan jumpa ini:

“I’m doing PCR to verify transgenic plants. First time I set up a PCR 3 samples with both positive and negative controls. It is pretty good! But the second time when I set up a large scale PCR with 50 samples, I got no PCR products at all ! including the positive control. I’m sure that the system is right, I didn’t miss any component. the mineral oil was added both time and the programs are same except hot lid was used the second time. I want to know does the hot lid increase the annealing temperature, thus ruin my PCR?”

 -suulei-

 

Dan antara jawapan yang buat saya berguling-guling ketawa [dalam hati]:

“There are 2 things to remember:-

(1) PCR stands for Polymerase Crap Reaction (i.e. it didn’t work because the moon was in the wrong phase, or you had the wrong coloured socks on etc…)

(2) There’s a car bumper sticker that goes – “Molecular biologists do it again and again and again and … …”

To summarise, there you probably didn’t do anything wrong, you just have to do it again. If it fails again then consider changing you stocks of dNTPs and primers – they’re usually the first to go.”

Hope this helps

zing

-zingmatter-

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