भारतीय विज्ञान शिक्षा एवं अनुसंधान संस्थान तिरुपति

INDIAN INSTITUTE OF SCIENCE EDUCATION AND RESEARCH TIRUPATI

(An Autonomous Institute under Ministry of Human Resource Development, Govt. of India)

Rami Reddy Nagar, Karakambadi Road, Mangalam P.O., Tirupati – 517 507
BIO-122

Set A

Experiment 1: Preparation of buffer

Each group to prepare the buffer (1X) for 50 ml:

Label the reagent bottle after preparing with Mon/Tue/Wed batch, Group No and Batch No. and place it on
the shelf of your seating place after informing the TA.

Calculate the concentration (xxxM) of the individual components of the buffer, after preparation.

TAE buffer
1X TAE buffer composition for 50 ml
pH 8.3
Tris base-(xxxxM)-0.484g
Glacial acetic acid (xxxM)-0.114 ml
EDTA- disodium salt-dihydrate (xxxM)- 0.0372 g
or 0.5 M stock EDTA disodium salt-dihydrate exists
from which how much volume of the solution need to
be be prepared to get desire concentration

References:

1. https://cshprotocols.cshlp.org/content/2006/1/pdb.rec8247

2. https://en.wikipedia.org/wiki/TAE_buffer

3. https://labchem-wako.fujifilm.com/asia/question/017743.html

4. https://cshprotocols.cshlp.org/content/2018/3/pdb.rec098756.full?text_only=true

5. https://www.aatbio.com/resources/buffer-preparations-and-recipes/sds-page-10x-sds-running-buffer
Experiment 2: Calibrating the micro-pipettes

Each of the students need measure the weight of the specified volume of the sample and plot a graph for
Volume of the sample vs Weight of the sample (in grams):

Sl . No. Volume range of the micropipette to be Volume of distilled Weight of the sample (in
used water to be pipetted grams)
(in µl) (as y-axis)
(as x-axis)
1 2

2 2 to 20 µl 5

3 10

4 20

5 20

6 50 Do not write anything

here
7 20 to 200 µl 100

8 150

9 200

10 100

11 200

12 350
100 to 1000 µl
13 500

14 750

15 1000

Calculate the density of the liquid given, from the plot.