Search found 22 matches

by siddharthsameer
Tue Oct 23, 2012 6:33 pm
Forum: Molecular Biology
Topic: molecular cloning failure
Replies: 3
Views: 2624

molecular cloning failure

Hello I am a master student and I am doing my master thesis in the cloning of keratinase sequence in competent cell.I tried several different methods and procedure for the cloning and I got the colonies but when I did the mini prep I had the good concentration of the DNA but when run on agarosei cou...
by siddharthsameer
Sat Aug 25, 2012 12:54 pm
Forum: Molecular Biology
Topic: isolation of plasmid (Midi prep)
Replies: 3
Views: 8105

Re: isolation of plasmid (Midi prep)

HI
THANKS FR YOUR REPLY... YES YOU ARE RIGHT, WHEN I DID THE ELECTROPHORESIS I DID NOT GET ANY RESULT OFMY DESIRED BAND, I DONT KNOW WHY IT HAPPENED ....AND WHAT SHOULD I DO NOW...KINDLY TELL ME
by siddharthsameer
Sat Aug 25, 2012 6:22 am
Forum: Molecular Biology
Topic: isolation of plasmid (Midi prep)
Replies: 3
Views: 8105

isolation of plasmid (Midi prep)

hello all I did my Midi and maxi prep and this time the concentration and mass were also good, but when I did the restriction digestion with the restriction enzyme to check my band I could not see the band at the desired position.. it is showing something else...M really tensed and I dont know wat t...
by siddharthsameer
Sun Aug 12, 2012 5:10 pm
Forum: Molecular Biology
Topic: Vector
Replies: 2
Views: 2540

Vector

hello all I would like to ask,is that i already of the used up my vector, and I plated the vector on one of the lb plate which i used as a negative control at the time of doing ligation,,So i would be trying to make the preculture from that and then the main culture and would do the miniprep to get ...
by siddharthsameer
Sun Aug 12, 2012 3:57 pm
Forum: Molecular Biology
Topic: pcr purification trouble
Replies: 4
Views: 5889

Re: pcr purification trouble

hi I measured the concentration through UV...so i am doing again PCR and this time I have decided to load all the sample for purification... Another thing I would like to ask,is that i already of the used up my vector, and I plated the vector on one of the lb plate which i used as a negative control...
by siddharthsameer
Sat Aug 11, 2012 8:10 am
Forum: Molecular Biology
Topic: pcr purification trouble
Replies: 4
Views: 5889

pcr purification trouble

Hello all I am into a trouble need some suggestion to correct myself. I had my pcr and I used 2 step pcr and i observed the band on agarose my desired sequence. Measured the concentration before and after pcr purification, but the concentration got down considerbaly. I did not purify the whole amoun...
by siddharthsameer
Wed Aug 01, 2012 6:54 pm
Forum: Molecular Biology
Topic: pcr purification
Replies: 2
Views: 1979

heelo i measured through UV spectro, but the concentration is too low,,,,do u know wats the reason
by siddharthsameer
Wed Aug 01, 2012 5:41 pm
Forum: Molecular Biology
Topic: pcr purification
Replies: 2
Views: 1979

pcr purification

Hello the concentration of my PCR product before purification was 1000microgram/ml and after the purification of my PCR product it has come down to 33.28 microgram/ ml.. what does it indicate.. is everyhing going fine or there is some problem in my pcr product.. kindly tell me....I am a big confusio...
by siddharthsameer
Wed Aug 01, 2012 4:17 pm
Forum: Molecular Biology
Topic: Restriction Digestion important
Replies: 1
Views: 1367

Restriction Digestion important

hello I need to ask one question and I am stuck with this kind;y help me... The concentration of my pcr product is 33.56microgram/ml and I want to make the restriction digestion .I want to have the 2 microgram of my product in the ristriction digestion volume of 50microlitre. How much volume should ...
by siddharthsameer
Wed Jul 25, 2012 8:49 am
Forum: Cell Biology
Topic: bacterial culture growth
Replies: 1
Views: 2659

bacterial culture growth

Hello everyone I have one question , why do we measure the ph of Preculture.I made a a preculture of e.coli tof 10 for the competent cell, and i was asked to measure the ph of the preculture, and I had 3 precultre in different shake falsk, and the following pH I measured . shake flask- pH 1 8,2 2 8,...
by siddharthsameer
Wed Jul 18, 2012 7:37 am
Forum: Molecular Biology
Topic: How to calculate mass of target Dna in ng
Replies: 1
Views: 2846

How to calculate mass of target Dna in ng

hello Everyone I have a small question, how do I calculate the mass of target DNA in ng as I am looking to calculate the amount of Restriction enzyme required for the digestion of my target DNA.Or How should I calculate the Volume needed for Restriction enzyme to digest my Vector And Target Dna. I h...
by siddharthsameer
Tue Jul 17, 2012 1:31 pm
Forum: Molecular Biology
Topic: restriction analysis
Replies: 14
Views: 12918

Re: restriction analysis

hello
I am about to start and my first step would be the amplification of my desired gene.. and then digestiona nd ligation into expression vector.. so If i start from the first step... thanks you so mucg for clearing my doubts...