Words, Words, Words - it's only words (Jul/21/2009 )
master
good for me........one more language I can learn...
mistress
@ div, I looked at that possibility, but i didnt have more than 5 ug from what i saw on the gel, then anywayz the Zymo kit should have worked...it had a capacity of 25 ug! Even that bombed...I checked the pH again eventhough i corrected the pH already
I normally end up with around 3ug from a 5 ug starting material..but this was around 600 ng!!!
distress
what bout buffers n all, e'thing working fine???? how do u do the final elution??? I use qiagen kit...trouble always used to lie in the buffers, if at all something used to go wrong....
damsel
i had that feeling and I made fresh buffers ! I still can't believe I had to do 12 replicate PCRs and and 3 cleanups to get 1.5 ug!!
mademoiselle
hell!!!
did u ask others in ur lab if they are hving any problem???? while using TE/MQ to elute out, maybe u can try warming it up a bit b4 using.....
chisel
they said they do have gr8 losses sometimes..I tried heating the EB for elution...still no luck..I tried all the basic troubleshooting that came to my mind, but nothing worked!!
I checked the pH, made the buffers fresh, left the columns for elution for 5 minutes, tried elution with warm EB, eluted it twice and thrice!! anything more ??
If this happens to my sequencing reactions, I'd be dead man!!
sizzle
what size fragments r u eluting..........some kits show problems wid larger sized DNA...
if e'one is having such losses, maybe its time to change the kit n get it from another company....
what volume do u elute in???
concentrating also doesn't help.......i mean after eluting wid MQ, n not EB??
sizzler
it was a 2 kb pcr product from a colony PCR...elution 12 ul x 2
Concentrating - why ? I dont have enough yield in total anyways!
I tried 2 companies and then it looks the same!!!
I've a meeting....will be back in an hour...have a nice day
drizzle
concen. coz i thought if u do elution in something like 50 ul, u conc it to 20 ul.........nyhow i dunno the purpose 4 which u require this dna....i need it for cloning, so conc. serves my purpose....
tried qiagen??? works good 4 us, so maybe.........
yeah bye....u too...
rain
what is happening here!!!! this is chit chat rooooooooom.. not a study centre!!!
and i got foood... seeemed as if i am from some place where i diont get food at all!!!