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Two-tier Multiple Choice Questions (MCQs) - How

effective are they: A pre-service teachers perspective

IOSTE - NW Europe, June 20-21 2011 Reading, UK,
Ms Alison Cullinane*, NCE-MSTL University of Limerick & Dr Maeve Liston, NCEMSTL Mary Immaculate College
Email: *

In recent times an increasing emphasis has been placed on the learning enhancement
purposes of assessment (Heywood, 2000; Holroyd, 2000). If changes are to occur in the
pedagogy of Biology and student learning, changes in assessment methods are needed
(Greer 2001; Brown et al. 1997; Gibbs 1999; BIO2010) as there is a very strong
relationship between assessment design and the effectiveness of student learning in the
biological sciences (Boud 1988; Mc Innis & Devlin 2002).
There is a general viewpoint that the methods in the Irish education system need to be
changed (Bennett & Kennedy, 2001; NCCA 2003; Looney, 2006). Assessment of Second
Level Biology in Ireland has come under increasing scrutiny in recent years. The Biology
Syllabus was altered in 2002, which included a change in the format and types of
questions used in the summative assessment examination paper. Many of the essay style
or extended answer questions were removed and replaced with short answer questions.
More recently a new proposed curriculum change was announced (NCCA, 2007).
Revision of this new syllabus will place an emphasis on the rejuvenation of the
summative examinations.
Traditionally the summative Biology examination paper in the Leaving Certificate
(established) programme is based on a paper and pencil design. It is argued such papers
largely assess lower order thinking, rarely asking question that assess higher cognitive
thinking (Bloom et al. 1956; Casserly 1984; Lord & Baviskar 2007). The current Biology
paper contains many closed questions requiring one word or one sentence answers. The
marking scheme of these papers are very restricting, only requiring key words and
terminology. There are few open questions which allow students to develop their answers
and demonstrate their understanding of a topic.
It has been found in many countries that there is a lack of formative assessment
techniques in many science classrooms even though numerous publications have
highlighted how formative assessment methods can significantly improve student
learning (Black & Wiliam 1998; Keeley 2008). As a prelude to this study, research was
undertaken to investigate the various types of formative assessment methods being
practiced in the Irish Science classroom. The findings highlighted the lack of assessment
for learning techniques, with a large proportion of class tests requiring rote learning and
the regurgitation of facts.
To combat this, an instrument called two-tier multiple choice questions (MCQs) were
developed for the Irish Biology classroom in the area of photosynthesis and

respiration. The main aim of this project was to design, develop and evaluate TwoTier Multiple Choice Questions on the topics of photosynthesis and respiration,
suitable for the Irish Senior Cycle Biology Syllabus. It was hoped that such
questions would add to the variety of formative assessment techniques which help
to promote higher thinking and identify misconception students may have in
Biology. The questions were completed and evaluated by Undergraduate Bachelor
of Science Education (in Biological Sciences) students at the University of
Limerick. This project also aims to investigate the Undergraduate B.Sc. (Ed)
students attitudes towards this formative assessment instrument.

The research questions were as follows:

1. Are Two-Tier MCQs an effective method of formative assessment for the Irish
Second Level Biology Classroom?
2. Can Two-Tier MCQs be used to identify misconceptions held by Irish students in
3. Can Two-Tier MCQs be used as a method to test students understanding of key
areas in Biology?
4. Would the Undergraduate pre-service Science teachers use the instrument as a
method of formative assessment in their classrooms in the future?
Two-tier Multiple Choice Questions (MCQs)
Two-tier MCQs have been used in the educational setting for many decades (Williams
2006), and are also referred to as Assertion-Reason Questions (Habeshaw et al. 1995;
Williams 2006) or Permutational Questions (Farthing et al. 1998; Bush 1999). Similar to
traditional MCQs, Two-Tier MCQs are classified into the genre of tests known as
Objective Tests. It is believed that this style of questioning first appeared in A-Level
examinations in the United Kingdom during the 1960s, however evidence has shown that
this format was used earlier than this in the United States medical examinations during
the 1950s (Moore, 1954). Despite this there is little academic research and literature
published on this long established method of assessment (Connelly 2004, Williams
Two-Tier MCQs are similar in format to traditional MCQs but as the name suggests, they
contain a second tier of questioning associated with the main question (see figure 1). The
aim of this second tier is to promote higher thinking and reasoning skills among students.
The first tier of the question usually pertains to a knowledge statement while the second
element of the question facilitates the testing of the students learning beyond recall and
into the higher levels of thinking (Haslam & Treagust 1987). Similar to traditional
MCQs, there are a key and distracters. The key is the correct answer and the distracter
describes the list of options present to distract from the key or the correct answer. The
design of Two-Tier MCQ uses Blooms taxonomy (1956) as a framework for promoting
different levels of thinking (Williams 2006). This instrument of questioning makes it
easier to test students higher level of thinking, more so than conventional MCQs
(Connelly 2004; Mann and Treagust 2000; Williams 2006). The purpose of the questions
is to help both student and teacher to identify student problem areas so they could be retaught to correct any misconceptions or areas of difficulty and develop a deeper

understanding of a topic (Mann & Treagust 1998; Odom, & Barrow 1995). Therefore a
Two-Tier MCQ instrument is a reliable and valid diagnostic measurement of second level
students understanding of biology (Haslam & Treagust 1987).

The Two-Tier MCQs was developed using guidelines that were employed by previous
researchers in this area, namely Mann & Treagust (1998), Haslam & Treagust (1987) and
Griffard and Wandersee (2001). This research project involved three phases:

Phase One
The first phase of this study involved the design and development of the Two-Tier
MCQs. The questions were developed specifically for the Irish Biology Senior Cycle
Higher Level Curriculum in the topics of photosynthesis and respiration. These topics
were chosen after an extensive literature review, which reported that students both
nationally and internationally had learning difficulties and held misconceptions in these
particular topics (Haslam & Treagust 1987; Griffard & Wandersee 2001; Liston 2009;
Mann & Treagust 1998; Storey 1989). The misconceptions identified by these
researchers were used in the development of the distracters for the second tier of these
questions. Thirteen questions were designed for the study (photosynthesis n=6 &
respiration n=7). Some questions were adapted from previous research, conducted on
Two-Tier MCQs, in particular Mann & Treagust (1998) and Haslam & Treagust (1987).

Phase Two
The questions were then evaluated by an academic member of the Life Science
Department at the University of Limerick. After this initial evaluation, the questions were
altered and deemed appropriate for the study.

Phase Three
The third phase of the study involved the evaluation of the questions developed by 1st
year and 3rd year Undergraduate BSc. (Ed.) (in Biological Sciences) Students at the
University of Limerick. This involved the completion of the questions. They were then
asked to fill out an evaluation form. The evaluation form sought their opinion on the
instrument as a method of formative assessment, as a method to diagnose misconceptions
and also their opinions on the suitability of using the instrument in second level Biology

Pre-service teachers were selected for this study from the Bachelor of Science Education
(B.Sc (Ed)) at the University of Limerick. Third level students were used for three
reasons; (i) gaining access to second level students in the time frame of the study was not
possible; (ii) The pre-service teachers would have recently completed Biology at Leaving
Certificate at second level which was a perquisite for taking part in the study. This would
provide a clear indication if secondary school students would benefit from the instrument;
(iii) to educate the pre-service teachers on different methods of formative assessment and
promoting assessment for learning in Biology and; (iv) the pre-service teachers would
give a good indication if practicing teachers would use the instrument in the second level

classroom as a method of formative assessment. The sample of students who volunteered

for the study (n=17) were from 1 st year (n=11) and 3rd year (n=6) of the degree
programme. The sample consisted of 58.8% females and 41.2% males.

This section provides an overview of the results obtained from the corrections of the
completed Two-Tier MCQs exercise and the pre-service teachers evaluation forms.

One of the main aims of Two-Tier MCQs is to identify misconceptions and
misunderstandings in science. On completion of the questions the researcher corrected
and recorded the undergraduate students answers. It is important to note that this study
was a voluntary study and the participants were high achieving students in Biology; many
receiving A grades in their Leaving Certificate examination. Despite this fact the
analysis of the Two-Tier MCQs exhibited some misconceptions. Table 1 and 2 below
illustrate the correct and incorrect student responses to the questions. As it can be seen
from the table below there were various incorrect responses to both the first and second
tiers of some questions. . The students obtained a mean score of 9.5/13. To be considered
correct both the first and second tier had to be answered accurately. Tables 1 and 2 below
highlights that even though the students answered the first tier correctly, they may have
answered the second tier incorrectly, which queries if they understood the theory behind
the topic; this is further highlighted in Table 3 below.
Table1. The correct and incorrect responses as given by the Undergraduate B.Sc. (Ed.)
Students to the Two-Tier MCQs on the topic of photosynthesis (N-17).
First Tier
Second Tier
Photosynthesis Correct
Incorrect Correct
Incorrect Total correct
Table 2. The correct and incorrect responses as given by the Undergraduate B.Sc. (Ed.)
Students to the Two-Tier MCQs on the topic of respiration (N=17).
First Tier
Second Tier
Correct Incorrect Correct Incorrect Total correct

Table 3 gives the number and percentage (in brackets) of the students correctly answering
both the first and second tiers of each question. The percentage of those correctly answering
only the first tier of the questions was also included to highlight the circumstances that
students were getting the right content answers without knowing/recognising the valid
reasons. A comparison of the percentage of students who correctly answered the knowledge
part of the questions with that of those who correctly answered both parts of the questions
suggests that many students may have learned facts without an adequate understanding of the
propositions and concepts involved (Peterson et al., 1989).

Table 3: The frequency of correct responses to the first tier compared with the frequency
of correct responses to both tiers.
No of Students Correctly Answering
First Tier
Both Tiers
17 (100%)
16 (94%)
17 (100%)
16 (94%)
17 (100%)
14 (82%)
17 (100%)
13 (76%)
12 (80%)
10 (59%)
15 (88%)
14 (82%)
16 (94%)
16 (94%)
17 (100%)
17 (100%)
11 (65%)
10 (59%)
16 (94%)
9 (53%)
12 (80%)
12 (80%)
10 (59%)
16 (94%)
8 (47%)
The tables above highlights the students held misconceptions in the various topics. There
was at least one wrong response to every question, except one, R2 which asked whether
respiration was a catabolic or anabolic reaction. In the second tier they were required to
establish a reason for their first selection and identify what a catabolic and an anabolic
reaction was. This question is similar to the question present in figure 1 on
photosynthesis. .
On comparing the correct answers from the 1 st and 3rd year students, the researcher
identified two misconceptions held by both groups in photosynthesis and three
misconceptions in respiration common to both years. The sections below will discuss
these five misconceptions in more detail:

Details of Identified Misconceptions

Minor misconceptions were observed from the questions on photosynthesis. Question
three (Figure 1.) showed that all of the students knew photosynthesis was an anabolic
reaction but the second tier of the question showed that 18% of the students did not
identify that the process of photosynthesis requires energy, to build up chemical
complexes, where it stores energy in high energy bonds and saves it for later. The
question is present in figure 1 below.

(Cullinane 2010)

Photosynthesis is a
A) Catabolic reaction
B) *Anabolic reaction
The reason for my answer is:

1) Photosynthesis is a catabolic reaction as the process involves the

oxidation of one molecule and the reduction of another allowing for
the break down of complex chemicals, which releases energy for the
plant to use.
2) *Photosynthesis is an anabolic reaction as the process of
photosynthesis requires energy to build up chemical complexes,
where it stores energy in high energy bonds and saved for later.
3) None of the above.
Figure 1: Two-tier MC questions developed on the topic of photosynthesis (Question three). *
Marks the correct answers to the second tier of the question. * The correct options in each tier

Question four showed that all of the students recognised water had an essential function
in the reaction of photosynthesis; however 24% of the students held misconceptions on
this topic. On analysis of the results some believed that the function of water in
photosynthesis was responsible for the transport for H+ ions in the light- independent
reaction. Therefore they were not able to reason out that, water supplies electrons in the
light dependent stage
Water has an essential function in the reaction of Photosynthesis:
A) *True
B) False
I know this because the function of water in Photosynthesis is to...
(1) Combine with Oxygen.
(2) Absorb light energy.
(3) Water is not essential and has no function in the process of
(4) Transport H+ ions in the light-independent (dark) reactions.
(5) Provide molecular oxygen for the light-independent (dark)
(6) *Supply electrons in the light-dependent reactions.
(Cullinane 2010)
Figure 2: Two-tier MC questions developed on the topic of photosynthesis Question Four). * The
correct options in each tier

Three misconceptions in respiration were identified to be common among both 1st and
3rd year students i.e. in questions 4, 6 and 7.
47% of respondents answered question four about respiration incorrectly. 53%
recognised that respiration took place in both plant and animal cells. But several students
(41%) selected number four to this question; Respiration took place in both animal and
plant cells but respiration only takes place during night time hours in plant cells. This
would indicate many of the students believed that respiration in plants only took place at
night when photosynthesis stopped.
Respiration occurs in both animal cells and plant cells.
A) *True
B) False
The reason for my answer is because:
1. Green plant cells respire when there is no light energy and carry out
the process of Photosynthesis when there is light energy. Animal cells
only respire in the day time.
2. Respiration only occurs in animal cells and not in plant cells. Plants
cells only Photosynthesis.
3. *Both plant and animal cells respire all the time. Plant cell respiration
takes place even during the day while Photosynthesis is occurring.
4. Respiration does take place in both animal and plant cells but
respiration only takes place during night time hours in plant cells

(Cullinane 2010)
Figure 3: Two-tier MC questions developed on the topic of respiration (Item four). * The correct
options in each tier

59% of respondents answered question six about respiration incorrectly. 7 out of the 17
did not recognise that glycolysis was an anaerobic process. 83% of those students
selected option 1 in the second tier, indicating they did not understand the full process of
glycolysis. That is, it is an anaerobic process and also they had the misconception that
carbon dioxide was released at this stage.
Glycolysis is the breakdown of glucose in aerobic conditions
A) True

B) *False

Reason for my answer is because:

1. Glycolysis can only take place in aerobic conditions; glucose is broken down
into 2 pyruvic acids and 2 ATP and carbon dioxide.
2. Glycolysis occurs in anaerobic conditions but glucose is not broken down, it is
the acetyl CoA which is broken down to pryruvic acid and 2 NAD+ molecules.
3. *Glycolysis is carried out in anaerobic conditions; where glucose is broken
into 2 pryruvic acids releasing 2 ATP molecules + water + 2 NADH molecules
4. Glycolysis is carried out in aerobic conditions; where glucose is broken into 2
pryruvic acids, releasing 2 ATP molecules + water + 2 NADH molecules

(Cullinane 2010)

Figure 4: Two-tier MC questions developed on the topic of respiration (Question Six). * The correct
options in each tier

53% of the students answered question seven on respiration incorrectly. All the students
with the exception of one, deduced that Acetyl CoA as the answer to the first tier of the

question. The misconception shown by both groups of students was that Acetyl CoA
enters the Krebs cycle and after one complete turn of the cycle, formed the products
phosphate, ADP, Hydrogen, oxygen and carbon dioxide). One of the most commonly
chosen responses by the 1st year group was number five (Glucose, carbon dioxide and
NAD+) indicating that they did not recognise that glucose was exhausted in respiration
and would not form an end product. No 3rd year student selected this as an answer.
In stage two of aerobic respiration (the Krebs cycle) the pyruvic acid enters the
mitochondrion of the cell where it loses carbon dioxide to form:
A) Acetyl CoB


B) Prvate

*D) Acetyl CoA

Reason for my answer is because ______________ goes on to enter the Krebs

cycle; after one complete turn of this cycle the below products are formed:
1. *Carbon dioxide, hydrogen, NADH and ATP
2. Oxygen and NADH and glycogen
3. ADP, carbon dioxide, glucose, cytosol,
4. Phosphate, ADP, hydrogen, oxygen and carbon dioxide
5. Glucose, carbon dioxide and NAD+

(Cullinane 2010)

Figure 5: Two-tier MCQs developed on the topic of respiration (Item six). * The correct options in
each tier

A summary of the common misconceptions the 1st and 3rd year B.Sc. (Ed) students held
on the topics of photosynthesis and respiration is illustrated below in table 3.
Table 3: Misconceptions on the topic of photosynthesis and respiration identified
within the sample of students in this study:

Students did not identify a anabolic reaction to be that the process of

photosynthesis requires energy, to build up chemical complexes, where it
stores energy in high energy bonds and saves it for later
Water has an essential function of transporting H + ions in the Lightindependent (Dark) stage


The students realised that respiration takes place in both animal and plant cells

but held the misconception that respiration only took place during night time
hours in plant cells.

The students failed to identify glycolysis as an anaerobic process and adding

to this they showed further lack of understanding of the process when they
considered CO2 was released during the process of glycolysis.
Most students were aware that it was Acetyl CoA that was formed when
pyruvic acid loses CO2 on entering the mitochondrion of the cell but many did
not identify the end products of the Krebs cycle to be CO 2, H, NADH and ATP.
The common misconception between both the 1st year and 3rd year groups was
that phosphate, ADP, H, O2 and CO2 were formed at the end of the Krebs
cycle. More alarmingly though 55% of the 1st year students believed glucose
CO2 and NAD+ were end products formed at the end of the cycle.

This section provides details of the feedback received from the Undergraduate B.Sc. (Ed)
students, on completion of the evaluation forms.
When asked on the evaluation form if they found the Two-Tier MCQs to be an effective
method of formative assessment, the results showed a very positive attitude towards the
instrument. 100% of the students found the questions to be an effective form of
assessment. When asked to elaborate on their answers, many stated that the instrument
would help the students to gain a better understanding of the topics. Some of their
comments were as follows:
Would help to give a better understanding
Make you fully understand the topic
very useful for revision
promote higher thinking and understanding
one is forced to think more about the question and topic
Question 2 of the evaluation form asked if the students would use the instrument as a
method of formative assessment in their future classrooms. 16 out of the 17 felt they
would use the instrument in their classroom.
Some of their comments were as follows:
They are a great way to challenge a class and maintain the
interest of eager learners
They also are a good assessment tool to see the levels of the
ability in a class
It would have helped me gain a more in-depth understanding
rather then simply learning off
They are quiet enjoyable to figure out It would help me
observe their understanding of the topic.

Many of the students felt they would use these types of questions because they were
very useful for revision. Indicating they would use them for formative assessment
purposes. Only one student selected that they would not use these types of questions for
future teaching. The student felt that he would like to teach the way (he) learned the
material But did go on the state he thought it was a quite a good concept.
The students were then asked if they thought the Two-Tier MCQs were an effective
method of assessment to diagnose misunderstanding and misconceptions that secondary
school students would have on a topic in biology. 100% responded positively expressing
they thought the questions were very good for this purpose.
Some of the comments were as follows:
Knowing someones reasoning is a good way to correct them if they are
making mistakes. It will make it easier to rectify the problem.
It justifies their reasons and beliefs for an answer which relates to the
core understanding of the material so these questions will highlight

This research project investigated the effectiveness of Two-Tier Multiple Choice
Questions as a method of formative assessment for the Irish Second Level Biology
Classroom. Third level students were chosen to evaluate the questions as these students
have completed the Leaving Certificate Biology Syllabus and received high grades in this
examination. The Two-Tier MCQs will investigate their knowledge but also their
understanding of different areas in photosynthesis and respiration.
From the comments and positive attitudes of the 1st and 3rd level undergraduate preservice Biology teachers that completed and evaluated the designed Two-Tier MCQs, it is
evident that there are many advantages to using these questions in the second level
biology classroom. The existence of a reliable and valid paper and pencil test instrument
which is easy to score and easily identify misconceptions of students is incredibly
valuable to teachers in their teaching and learning. It was suggested that these questions
would contribute greatly to formative assessment exercises, promoting assessment for
learning and a deeper level of thinking. The students were required to use higher levels of
thinking in order to reason out their answers for the second tier. The questions were also a
suitable method to identify misconceptions held by Irish students in Biology and to test
students understanding of key areas in Biology. The results highlighted that the students
held many misconceptions in the topics of respiration and photosynthesis. Teachers could
use this instrument as a diagnostic test, to investigate areas of misunderstanding among
students. This research identifies areas in the topics of photosynthesis and respiration
that second level students may find difficult to understand leading to misconceptions.
These questions could also be used in professional development lectures of pre-service
science teachers and continuous professional development of qualified teachers.
The findings have highlighted further research that needs to be carried out in this area:

As the questions are intended for the secondary school classroom, to add to this study
practicing teachers in Irish classrooms will be asked to evaluate the two-tier MCQs
and give their opinion on the instrument as a method of formative assessment in
biology on the topics of photosynthesis and respiration.

Adding a confidence rating at the end of the questions in the future would further
identify if the students were guessing or understood the answer provided.

The questions designed here were more suitable for higher level students; future work
would entail the design and evaluation of two-tier questions for weaker ability or
ordinary level students.


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